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Generation of recombinant antibodies

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Abstract

Recombinant antibody technology is opening new perspectives for the development of novel therapeutic and diagnostic agents. In this review we focus on advances in the generation of both genetically engineered humanized and fully human monoclonal antibodies. Methods for their production in different expression systems are also discussed.

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References

  1. Riethmüller, G., Schneider Gadicke, E., and Johnson, J. P. (1993) Monoclonal antibodies in cancer therapy. Curr. Opin. Immunol. 5, 732–739.

    PubMed  Google Scholar 

  2. Jaffers, G. J., Fuller, T. C., Cosimi, A. B., Russell, P. S., Winn, H. J., and Colvin, R. B. (1986) Monoclonal antibody therapy. Anti-idiotypic and non-anti-idiotypic antibodies to OKT3 arising despite intense immunosuppression. Transplantation 41, 572–578.

    PubMed  CAS  Google Scholar 

  3. Khazaeli, M. B., Conry, R. M., and LoBuglio, A. F. (1994) Human immune response to monoclonal antibodies. J. Immunotherapy 15, 42–52.

    Article  CAS  Google Scholar 

  4. James, K. (1994) Human monoclonal antibody technology, in Handbook of Experimental Pharmacology, Vol. 113: The Pharmacology of Monoclonal Antibodies (Rosenberg, M., and Moore, G. P., eds.), Springer-Verlag, Berlin, Heidelberg, pp. 3–22.

    Google Scholar 

  5. Bird, R. E., Hardman, K. D., Jacobson, J. W., Johnson, S., Kaufman, B. M., Lee, S.-M., Lee, T., Pope, S. H., Riordan, G. S., and Whitlow, M. (1988) Single-chain antigen-binding proteins. Science 242, 423–426.

    PubMed  CAS  Google Scholar 

  6. Huston, J. S., Levinson, D., Mudgett Hunter, M., Tai, M. S., Novotny, J., Margolies, M. N., Ridge, R. J., Bruccoleri, R. E., Haber, E., Crea, R., and Oppermann, H. (1988) Protein engineering of anti-body binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc. Natl. Acad. Sci. USA 85, 5879–5883.

    PubMed  CAS  Google Scholar 

  7. Borrebaeck, C. A., Danielsson, L., and Moller, S. A. (1988) Human monoclonal antibodies produced by primary in vitro immunization of peripheral blood lymphocytes. Proc. Natl. Acad. Sci. USA 85, 3995–3999.

    PubMed  CAS  Google Scholar 

  8. Brüggemann, M., Spicer, C., Buluwela, L., Rosewell, I., Barton, S., Surani, M. A., and Rabbitts, T. H. (1991) Human antibody production in transgenic mice: expression from 100 kb of the human IgH locus. Eur. J. Immunol. 21, 1323–1326.

    PubMed  Google Scholar 

  9. Lonberg, N., Taylor, L. D., Harding, F. A., Trounstine, M., Higgins, K. M., Schramm, S. R., Kuo, C. C., Mashayekh, R., Wymore, K., McCabe, J. G., and et al. (1994) Antigen-specific human antibodies from mice comprising four distinct genetic modifications. Nature 368, 856–859.

    PubMed  CAS  Google Scholar 

  10. Lonberg, N., and Huszar, D. (1995) Human antibodies from transgenic mice. Int. Rev. Immunol. 13, 65–93.

    PubMed  CAS  Google Scholar 

  11. Jakobovits, A. (1995) Production of fully human antibodies by transgenic mice. Curr. Opin. Biotechnol. 6, 561–566.

    PubMed  CAS  Google Scholar 

  12. Marks, J. D., Hoogenboom, H. R., Bonnert, T. P., McCafferty, J., Griffiths, A. D., and Winter, G. (1991) By-passing immunization. Human antibodies from V-gene libraries displayed on phage. J. Mol. Biol. 222, 581–597.

    PubMed  CAS  Google Scholar 

  13. Welschof, M., Terness, P., Kolbinger, F., Zewe, M., Dübel, S., Dörsam, H., Hain, C., Finger, M., Jung, M., Moldenhauer, G., Hayashi, N., Little, M., and Opelz, G. (1995) Amino acid sequence based PCR primers for amplification of rearranged human heavy and light chain immunoglobulin variable region genes. J. Immunol. Methods 179, 203–214.

    PubMed  CAS  Google Scholar 

  14. Lagerkvist, A. C., Furebring, C., and Borrebaeck, C. A. (1995) Single, antigen-specific B cells used to generate Fab fragments using CD40-mediated amplification or direct PCR cloning. Biotechniques 18, 862–869.

    PubMed  CAS  Google Scholar 

  15. Terness, P., Welschof, M., Moldenhauer, G., Jung, M., Moroder, L., Kirchhoff, F., Kipriyanov, S., Little, M., and Opelz, G. (1997) Idiotypic vaccine for treatment of human B-cell lymphoma. Construction of IgG variable regions from single malignant B cells. Hum. Immunol. 56, 17–27.

    PubMed  CAS  Google Scholar 

  16. Liu, A. Y., Robinson, R. R., Hellström, K. E., Murray, E. D., Chang, C. P., and Hellström, I. (1987) Chimeric mouse-human IgG1 antibody that can mediate lysis of cancer cells. Proc. Natl. Acad. Sci. USA 84, 3439–3443.

    PubMed  CAS  Google Scholar 

  17. Padlan, E. A. (1994) Anatomy of the antibody molecule. Mol. Immunol. 31, 169–217.

    PubMed  CAS  Google Scholar 

  18. Jones, P. T., Dear, P. H., Foote, J., Neuberger, M. S., and Winter, G. (1986) Replacing the complementarity-determing regions in a human antibody with those from a mouse. Nature 321, 522–525.

    PubMed  CAS  Google Scholar 

  19. Verhoeyen, M., Milstein, C., and Winter, G. (1988) Reshaping human antibodies: grafting an antilysozyme activity. Science 239, 1534–1536.

    PubMed  CAS  Google Scholar 

  20. Queen, C., Schneider, W. P., Selick, H. E., Payne, P. W., Landolfi, N. F., Duncan, J. F., Avdalovic, N. M., Levitt, M., Junghans, R. P., and Waldmann, T. A. (1989) A humanized antibody that binds to the interleukin 2 receptor. Proc. Natl. Acad. Sci. USA 86, 10,029–10,033.

    CAS  Google Scholar 

  21. Co, M. S., and Queen, C. (1991) Humanized antibodies for therapy. Nature 351, 501,502.

    Google Scholar 

  22. Ohtomo, T., Tsuchiya, M., Sato, K., Shimizu, K., Moriuchi, S., Miyao, Y., Akimoto, T., Akamatsu, K., Hayakawa, T., and Ohsugi, Y. (1995) Humanization of mouse ONS-M21 antibody with the aid of hybrid variable regions. Mol. Immunol. 32, 407–416.

    PubMed  CAS  Google Scholar 

  23. Kolbinger, F., Saldanha, J., Hardman, N., and Bendig, M. M. (1993) Humanization of a mouse anti-human IgE antibody: a potential therapeutic for IgE-mediated allergies. Protein Eng. 6, 971–980.

    PubMed  CAS  Google Scholar 

  24. Roguska, M. A., Pedersen, J. T., Henry, A. H., Searle, S. M., Roja, C. M., Avery, B., Hoffee, M., Cook, S., Lambert, J. M., Blattler, W. A., Rees, A. R., and Guild, B. C. (1996) A comparison of two murine monoclonal antibodies humanized by CDR-grafting and variable domain resurfacing. Protein Eng. 9, 895–904.

    Google Scholar 

  25. Kettleborough, C. A., Saldanha, J., Heath, V. J., Morrison, C. J., and Bendig, M. M. (1991) Humanization of a mouse monoclonal antibody of CDR-grafting: the importance of framework residues on loop conformation. Protein Eng. 4, 773–783.

    PubMed  CAS  Google Scholar 

  26. Mateo, C., Moreno, E., Amour, K., Lombardero, J., Harris, W., and Perez, R. (1997) Humanization of a mouse monoclonal antibody that blocks the epidermal growth factor receptor: recovery of antagonistic activity. Immunotechnology 3, 71–81.

    PubMed  CAS  Google Scholar 

  27. Carter, P., Presta, L., Gorman, C. M., Ridgway, J. B., Henner, D., Wong, W. L., Rowland, A. M., Kotts, C., Carver, M. E., and Shepard, H. M. (1992) Humanization of an anti-p185HER2 antibody for human cancer therapy. Proc. Natl. Acad. Sci. USA 89, 4285–4289.

    PubMed  CAS  Google Scholar 

  28. Kashmiri, S. V., Shu, L., Padlan, E. A., Milenic, D. E., Schlom, J., and Hand, P. H. (1995) Generation, characterization, and in vivo studies of humanized anti-carcinoma antibody CC49. Hybridoma 14, 461–473.

    Article  PubMed  CAS  Google Scholar 

  29. Shalaby, M. R., Shepard, H. M., Presta, L., Rodrigues, M. L., Beverley, P. C., Feldmann, M., and Carter, P. (1992) Development of humanized bispecific antibodies reactive with cytotoxic lymphocytes and tumor cells overexpressing the HER2 protooncogene. J. Exp. Med. 175, 217–225.

    PubMed  CAS  Google Scholar 

  30. Rodrigues, M. L., Shalaby, M. R., Werther, W., Presta, L., and Carter, P. (1992) Engineering a humanized bispecific F(ab’)2 fragment for improved binding to T cells. Int. J. Cancer (Suppl.) 7, 45–50.

    CAS  Google Scholar 

  31. Adair, J. R., Athwal, D. S., Bodmer, M., Bright, S. M., Collins, A., Pulito, V. L., Rao, P. E., Reedman, R., Rothermel, A. L., Xu, D., Zivin, R. A., and Jolliffe, L. K. (1994) Humanization of the murine anti-human CD3 monoclonal antibody OKT3. Hum. Antibod. Hybridomas 5, 41–47.

    CAS  Google Scholar 

  32. Pulito, V. L., Roberts, V. A., Adair, J. R., Rothermel, A. L., Collins, A. M., Varga, S. S., Martocello, C., Bodmer, M. W., Jolliffe, L. K., and Zivin, R. A. (1996) Humanization and molecular modeling of the anti-CD4 monoclonal antibody, OKT4A. J. Immunol. 156, 2840–2850.

    Google Scholar 

  33. Hsiao, K.-C., Bajorath, J., and Harris, L. J. (1994) Humanization of 60.3, an anti-CD18 antibody; importance of the L2 loop. Protein Eng. 7, 815–822.

    PubMed  CAS  Google Scholar 

  34. Leung, S.-O., Goldenberg, D. M., Dion, A., Pellegrini, M. C., Shevitz, J., Shih, L. B., and Hansen, H. J. (1995) Construction and characterization of a humanized, internalizing, B-cell (CD22)-specific, leukemia/lymphoma antibody, LL2. Mol. Immunol. 32, 1413–1427.

    PubMed  CAS  Google Scholar 

  35. Graziano, R. F., Tempest, P. R., White, P., Keler, T., Deo, Y., Ghebremariam, H., Coleman, K., Pfefferkorn, L. C., Fanger, M. W., and Guyre, P. M. (1995) Construction and characterization of a humanized anti-gamma-Ig receptor type I (Fc gamma RI) monoclonal antibody. J. Immunol. 155, 4996–5002.

    PubMed  CAS  Google Scholar 

  36. Riechmann, L., Clark, M., Waldmann, H., and Winter, G. (1988) Reshaping human antibodies for therapy. Nature 332, 323–327.

    PubMed  CAS  Google Scholar 

  37. Tempest, P. R., Bremner, P., Lambert, M., Taylor, G., Furze, J. M., Carr, F. J., and Harris, W. J. (1991) Reshaping a human monoclonal antibody to inhibit human respiratory syncytical virus infection in vivo. Biol Technology 9, 266–271.

    CAS  Google Scholar 

  38. Padlan, E. A. (1991) A possible procedure for reducing the immunogenicity of antibody variable domains while preserving their ligand-binding properties. Mol. Immunol. 28, 489–498.

    PubMed  CAS  Google Scholar 

  39. Pedersen, J. T., Henry, A. H., Searle, S. J., Guild, B. C., Roguska, M., and Rees, A. R. (1994) Comparison of surface accessible residues in human and murine immunoglobulin Fv domains. Implication for humanization of murine antibodies. J. Mol. Biol. 235, 959–973.

    PubMed  CAS  Google Scholar 

  40. Mark, G. E., and Padlan, E. A. (1994) Humanization of monoclonal antibodies, in Handbook of Experimental Pharmacology, Vol. 113: The Pharmacology of Monoclonal Antibodies (Rosenberg, M., and Moore, G. P., eds.), Springer-Verlag, Berlin, Heidelberg, pp. 105–134.

    Google Scholar 

  41. Roguska, M. A., Pedersen, J. T., Keddy, C. A., Henry, A. H., Searle, S. J., Lambert, J. M., Goldmacher, V. S., Blattler, W. A., Rees, A. R., and Guild, B. C. (1994) Humanization of murine monoclonal antibodies through variable domain resurfacing. Proc. Natl. Acad. Sci. USA 91, 969–973.

    PubMed  CAS  Google Scholar 

  42. Woodle, E. S., Thistlethwaite, J. R., Jolliffe, L. K., Zivin, R. A., Collins, A., Adair, J. R., Bodmer, M., Athwal, D., Alegre, M.-L., and Bluestone, J. A. (1992) Humanized OKT3 antibodies: successful transfer of immune modulating properties and idiotype expression. J. Immunol. 148, 2756–2763.

    PubMed  CAS  Google Scholar 

  43. Alegre, M.-L., Collins, A., Pulito, V. L., Brosius, R. A., Olson, W. C., Zivin, R. A., Knowles, R., Thistlethwaite, J. R., Jolliffe, L. K., and Bluestone, J. A. (1992) Effect of a single amino acid mutation on the activating and immunosuppressive properties of a “humanized” OKT3 monoclonal antibody. J. Immunol. 148, 3461–3468.

    PubMed  CAS  Google Scholar 

  44. Alegre, M.-L., Peterson, L. J., Xu, D., Sattar, H. A., Jeyarajah, D. R., Knowlakowski, K., Thistlethwaite, J. R., Zivin, R. A., Jolliffe, L. K., and Bluestone, J. A. (1994) A non-activating “humanized” anti-CD3 monoclonal antibody retains immunosuppressive properties in vivo. Transplantation 57, 1537–1543.

    PubMed  CAS  Google Scholar 

  45. Sun, L. K., Curtis, P., Rakowicz-Szulczynska, E., Ghrayeb, J., Chang, N., Morrison, S. L., and Koprowski, H. (1987) Chimeric antibody with human constant regions and mouse variable regins directed against carcinoma-associated antigen 17-1A. Proc. Natl. Acad. Sci. USA 84, 214–218.

    PubMed  CAS  Google Scholar 

  46. Bebbington, C. R., Renner, G., Thomspon, S., King, D., Abrams, D., and Yarranton, G. T. (1992) High-level expression of a recombinant antibody from myeloma cells using a glutamine synthetase gene as an amplifiable selectable marker. Bio/Technology 10, 169–175.

    PubMed  CAS  Google Scholar 

  47. Page, M. J., and Sydenham, M. A. (1991) High level expression of the humanized monoclonal antibody CAMPATH-1H in Chinese hamster ovary cells. Bio/Technology 9, 64–68.

    PubMed  CAS  Google Scholar 

  48. Fouser, L. A., Swanberg, S. L., Lin, B.-Y., Benedict, M., Kelleher, K., Cumming, D. A., and Riedel, G. E. (1992) High level expression of a chimeric anti-ganglioside GD2 antibody: genomic kappa sequences improve expression in COS and CHO cells. Bio/Technology 10, 1121–1127.

    PubMed  CAS  Google Scholar 

  49. Dorai, H., and Moore, G. P. (1987) The effect of dihydrofolate reductase-mediated gene amplification on the expression of transfected immunoglobulin genes. J. Immunol. 139, 4232–4241.

    PubMed  CAS  Google Scholar 

  50. Shitara, K., Nakamura, K., Tokutake-Tanaka, Y., Fukushima, M., and Hanai, N. (1994) A new vector for the high level expression of chimeric antibodies in myeloma cells. J. Immunol. Methods 167, 271–278.

    PubMed  CAS  Google Scholar 

  51. Shu, L., Qi, C.-F., Schlom, J., and Kashmiri, S. V. S. (1993) Secretion of a single-gene-encoded immunoglobulin from myeloma cells. Proc. Natl. Acad. Sci. USA 90, 7995–7999.

    PubMed  CAS  Google Scholar 

  52. Smith, J. W., Hu, D., Satterthwait, A., Pinzsweeney, S., and Barbas, C. F. (1994) Building synthetic anti-bidies as adhesive ligands for integrins. J. Biol. Chem. 269, 32,788–32,795.

    CAS  Google Scholar 

  53. Traunecker, A., Lanzavecchia, A., and Karjalainen, K. (1991) Bispecific single chain molecules (Janusins) target cytotoxic lymphocytes on HIV infected cells. EMBO J. 10, 3655–3659.

    PubMed  CAS  Google Scholar 

  54. Traunecker, A., Lanzavecchia, A., and Karjalainen, K. (1992) Janusin: new molecular design for bispecific reagents. Int. J. Cancer (Suppl.) 7, 51,52.

    Google Scholar 

  55. Chamow, S. M., Zhang, D. Z., Tan, X. Y., Mhatre, S. M., Marsters, S. A., Peers, D. H., Byrn, R. A., Ashkenazi, A., and Junghans, R. P. (1994) A humanized, bispecific immunoadhesin-antibody that retargets CD3+ effectors to kill HIV-1-infected cells. J. Immunol. 153, 4268–4280.

    PubMed  CAS  Google Scholar 

  56. Ridgway, J. B., Presta, L. G., and Carter, P. (1996) ‚Knobs-into-holes‘ engineering of antibody CH3 domains for heavy chain heterodimerization. Protein Eng. 9, 617–621.

    Google Scholar 

  57. De Sutter, K., Feys, V., Van de Voorde, A., and Fiers, W. (1992) Production of functionally active murine and murine::human chimeric F(ab’)2 fragments in COS-1 cells. Gene 113, 223–230.

    PubMed  Google Scholar 

  58. Young, R. J., Owens, R. J., Mackay, G. A., Chan, C. M. W., Shi, J., Hide, M., Francis, D. M., Henry, A. J., Sutton, B. J., and Gould, H. J. (1995) Secretion of recombinant human IgE-Fc by mammalian cells and biological activity of glycosilation site mutants. Protein Eng. 8, 193–199.

    PubMed  CAS  Google Scholar 

  59. King, D. J., Byron, O. D., Mountain, A., Weir, N., Harvey, A., Lawson, A. D. G., Proudfoot, K. A., Baldock, D., Harding, S. E., Yarranton, G. T., and Owens, R. J. (1993) Expression, purification and characterization of B72.3 Fv fragments. Biochem. J. 290, 723–729.

    PubMed  CAS  Google Scholar 

  60. Jost, C. R., Kurucz, I., Jacobus, C. M., Titus, J. A., George, A. J., and Segal, D. M. (1994) Mammalian expression and secretion of functional single-chain Fv molecules. J. Biol. Chem. 269, 26,267–26,273.

    CAS  Google Scholar 

  61. Hu, S., Shively, L., Raubitschek, A., Sherman, M., Williams, L. E., Wong, J. Y., Shively, J. E., and Wu, A. M. (1996) Minibody: A novel engineered anti-carcinoembryonic antigen antibody fragment (single-chain Fv-CH3) which exhibits rapid, high-level targeting of xenografts. Cancer Res. 56, 3055–3061.

    Google Scholar 

  62. Libyh, M. T., Goossens, D., Oudin, S., Gupta, N., Dervillez, X., Juszczak, G., Cornillet, P., Bougy, F., Reveil, B., Philbert, F., Tabary, T., Klatzmann, D., Rouger, P., and Cohen, J. H. (1997) A recombinant human scFv anti-Rh(D) antibody with multiple valences using a C-terminal fragment of C4-binding protein. Blood 90, 3978–3983.

    PubMed  CAS  Google Scholar 

  63. Gillies, S. D., Lan, Y., Wesolowski, J. S., Qian, X., Reisfeld, R. A., Holden, S., Super, M., and Lo, K.-M. (1998) Antibody-IL-12 fusion proteins are effective in SCID mouse models of prostate and colon carcinoma matastases. J. Immunol. 160, 6195–6203.

    PubMed  CAS  Google Scholar 

  64. Dorai, H., McCartney, J. E., Hudziak, R. M., Tai, M.-S., Laminet, A. A., Houston, L. L., Huston, J. S., and Oppermann, H. (1994) Mammalian cell expression of single-chain Fv (sFv) antibody proteins and their C-terminal fusions with interleukin-2 and other effector domains. Bio/Technology 12, 890–897.

    PubMed  CAS  Google Scholar 

  65. Jost, C. R., Titus, J. A., Kurucz, I., and Segal, D. M. (1996) A single-chain bispecific Fv2 molecule produced in mammalian cells redirects lysis by activated CTL. Mol. Immunol. 33, 211–219.

    Google Scholar 

  66. Mack, M., Riethmüller, G., and Kufer, P. (1995) A small bispecific antibody construct expressed as a functional single-chain molecule with high tumor cell cytotoxicity. Proc. Natl. Acad. Sci. USA 92, 7021–7025.

    PubMed  CAS  Google Scholar 

  67. Limonta, J., Pedraza, A., Rodriguez, A., Freyre, F. M., Barral, A. M., Castro, F. O., Lleonart, R., Gracia, C. A., Gavilondo, J. V., and de la Fuente, J. (1995) Production of active anti-CD6 mouse/human chimeric antibodies in the milk of transgenic mice. Immunotechnology 1, 107–113.

    PubMed  CAS  Google Scholar 

  68. Richardson, J. H., Sodroski, J. G., Waldmann, T. A., and Marasco, W. A. (1995) Phenotypic knockout of the high-affinity human interleukin 2 receptor by intracellular single-chain antibodies against the a subunit of the receptor. Proc. Natl. Acad. Sci. USA 92, 3137–3141.

    PubMed  CAS  Google Scholar 

  69. Biocca, S., Pierandrei-Amaldi, P., Campioni, N., and Cattaneo, A. (1994) Intracellular immunization with cytosolic recombinant antibodies. Bio/Technology 12, 396–399.

    PubMed  CAS  Google Scholar 

  70. Biocca, S., Ruberti, F., Tafani, M., Pierandrei-Amaldi, P., and Cattaneo, A. (1995) Redox state of single chain Fv fragments targeted to the endoplasmic reticulum, cytosol and mitochondria. Bio/Technology 13, 1110–1115.

    PubMed  CAS  Google Scholar 

  71. Jiang, W., Venugopal, K., and Gould, E. A. (1995) Intracellular interference of tick-borne flavivirus infection by using a single-chain antibody fragment delivered by recombinant Sindbis virus. J. Virol. 69, 1044–1049.

    PubMed  CAS  Google Scholar 

  72. Levin, R., Mhashilkar, A. M., Dorfman, T., Bukovsky, A., Zani, C., Bagley, J., Hinkula, J., Niedrig, M., Albert, J., Wahren, B., Gottlinger, H. G., and Marasco, W. A. (1997) Inhibition of early and late events of the HIV-1 replication cycle by cytoplasmic Fab intrabodies against the matrix protein, p17. Mol. Med. 3, 96–110.

    PubMed  CAS  Google Scholar 

  73. Mhashilkar, A. M., Biswas, D. K., LaVecchio, J., Pardee, A. B., and Marasco, W. A. (1997) Inhibition of human immunodeficiency virus type 1 replication in vitro by a novel combination of anti-Tat single-chain intrabodies and NF-kappa B antagonists. J. Virol. 71, 6486–6494.

    PubMed  CAS  Google Scholar 

  74. Rode, H.-J., Little, M., Fuchs, P., Dorsam, H., Schooltink, H., de Ines, C., Dubel, S., and Breitling, F. (1996) Cell surface display of a single-chain antibody for attaching polypeptides. Biotechniques 21, 650–658.

    Google Scholar 

  75. Eshhar, Z., Bach, N., Fitzer Attas, C. J., Gross, G., Lustgarten, J., Waks, T., and Schindler, D. G. (1996) The T-body approach: potential for cancer immunotherapy. Springer Semin. Immunopathol. 18, 199–209.

    Google Scholar 

  76. Eshhar, Z. (1997) Tumor-specific T-bodies: towards clinical application. Cancer Immunol. Immunother. 45, 131–136.

    PubMed  CAS  Google Scholar 

  77. Eshhar, Z., Waks, T., Gross, G., and Schindler, D. G. (1993) Specific activation and targeting of cytotoxic lymphocytes through chimeric single chains consisting of antibody-binding domains and the gamma or zeta subunits of the immunoglobulin and T-cell receptors. Proc. Natl. Acad. Sci. USA 90, 720–724.

    PubMed  CAS  Google Scholar 

  78. Hwu, P., Shafer, G. E., Treisman, J., Schindler, D. G., Gross, G., Cowherd, R., Rosenberg, S. A., and Eshhar, Z. (1993) Lysis of ovarian cancer cells by human lymphocytes redirected with a chimeric gene composed of an antibody variable region and the Fc receptor gamma chain. J. Exp. Med. 178, 361–366.

    PubMed  CAS  Google Scholar 

  79. Poul, M.-A., Cerutti, M., Chaabihi, H., Ticchioni, M., Deramoudt, F.-X., Bernard, A., Devauchelle, G., Kaczorek, M., and Lefranc, M.-P. (1995) Cassette baculovirus vectors for the production of chimeric, humanized, or human antibodies in insect cells. Eur. J. Immunol. 25, 2005–2009.

    PubMed  CAS  Google Scholar 

  80. Kretzschmar, T., Aoustin, L., Zingel, O., Marangi, M., Vonach, B., Towbin, H., and Geiser, M. (1996) High-level expression in insect cells and purification of secreted monomeric single-chain Fv antibodies. J. Immunol. Methods 195, 93–101.

    Google Scholar 

  81. Brocks, B., Rode, H.-J., Klein, M., Gerlach, E., Dübel, S., Little, M., Pfitzenmaier, K., and Moosmayer, D. (1997) A TNF receptor antagonistic scFv, which is not secreted in mammalian cells, is expressed as a soluble mono-and bivalent scFv derivative in insect cells. Immunotechnology 3, 173–184.

    PubMed  CAS  Google Scholar 

  82. Ma, J. K., Hiatt, A., Hein, M., Vine, N. D., Wang, F., Stabila, P., van Dolleweerd, C., Mostov, K., and Lehner, T. (1995) Generation and assembly of secretory antibodies in plants. Science 268, 716–719.

    PubMed  CAS  Google Scholar 

  83. Ma, J. K., and Hein, M. B. (1995) Immunotherapeutic potential of antibodies produced in plants. Trends Biotechnol. 13, 522–527.

    PubMed  CAS  Google Scholar 

  84. Hiatt, A. C., Cafferkey, R., and Bowdish, K. (1989) Production of antibodies in transgenic plants. Nature 342, 76–78.

    PubMed  CAS  Google Scholar 

  85. During, K., Hippe, S., Kreuzaler, E., and Schell, J. (1990) Synthesis and self-assembly of a functional antibody in transgenic Nicotiana tabacum. Plant Mol. Biol. 15, 281–293.

    PubMed  CAS  Google Scholar 

  86. Tavladoraki, P., Benvenuto, E., Trinca, S., De Martinis, D., Cattaneo, A., and Galeffi, P. (1993) Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. Nature 366, 469–472.

    PubMed  CAS  Google Scholar 

  87. Fiedler, U., and Conrad, U. (1995) High-level production and long-term storage of engineered antibodies in transgenic tabacco seeds. Bio/Technology 13, 1090–1093.

    PubMed  CAS  Google Scholar 

  88. Wood, C. R., Boss, M. A., Kenten, J. H., Calvert, J. E., Roberts, N. A., and Emtage, J. S. (1985) The synthesis and in vivo assembly of functional antibodies in yeast. Nature 314, 446–449.

    PubMed  CAS  Google Scholar 

  89. Horwitz, A. H., Chang, C. P., Better, M., Hellstrom, K. E., and Robinson, R. R. (1988) Secretion of functional antibody and Fab fragment from yeast cells. Proc. Natl. Acad. Sci. USA 85, 8678–8682.

    PubMed  CAS  Google Scholar 

  90. Boder, E. T., and Wittrup, K. D. (1997) Yeast surface display for screening combinatorial polypeptide libraries. Nature Biotechnol. 15, 553–557.

    CAS  Google Scholar 

  91. Cregg, J. M., Vedvick, T. S., and Raschke, W. C. (1993) Recent advances in the expression of foreign genes in Pichia pastoris. Bio/Technology 11, 905–910.

    PubMed  CAS  Google Scholar 

  92. Hollenberg, C. P., and Gelissen, G. (1997) Production of recombinant proteins by methylotrophic yeasts. Curr. Opin. Biotechnol. 8, 554–560.

    PubMed  CAS  Google Scholar 

  93. Ridder, R., Schmitz, R., Legay, F., and Gram, H. (1995) Generation of rabbit monoclonal antibody fragments from a combinatorial phage display library and their production in the yeast Pichia pastoris. Bio/Technology 13, 255–260.

    PubMed  CAS  Google Scholar 

  94. Eldin, P., Pauza, M. E., Hieda, Y., Lin, G., Murtaugh, M. P., Pentel, P. R., and Pennell, C. A. (1997) High-level secretion of two antibody single chain Fv fragments by Pichia pastoris. J. Immunol. Methods 201, 67–75.

    PubMed  CAS  Google Scholar 

  95. FitzGerald, K., Holliger, P., and Winter, G. (1997) Improved tumour targeting by disulphide stabilized diabodies expressed in Pichia pastoris. Protein Eng. 10, 1221–1225.

    PubMed  CAS  Google Scholar 

  96. Nyyssönen, E., Penttilä, M., Harkki, A., Saloheimo, A., Knowles, J. K. C., and Keränen, S. (1993) Efficient production of antibody fragments by the filamentous fungus Trichoderma reesei. Bio/Technology 11, 591–595.

    PubMed  Google Scholar 

  97. Boss, M. A., Kenten, J. H., Wood, C. R., and Emtage, J. S. (1984) Assembly of functional antibodies from immunoglobulin heavy and light chains synthesized in E. coli. Nucl. Acids Res. 12, 3791–3806.

    PubMed  CAS  Google Scholar 

  98. Wu, X.-C., Ng, S.-C., Near, R. I., and Wong, S.-L. (1993) Efficient production of a functional single-chain antidigoxin antibody via an engineered Bacillus subtilis expression-secretion system. Bio/Technology 11, 71–76.

    PubMed  CAS  Google Scholar 

  99. Better, M., Cheng, C. P., Robinson, R. R., and Horowitz, A. H. (1988) E. coli secretion of an active chimeric antibody fragment. Science 240, 1041–1043.

    PubMed  CAS  Google Scholar 

  100. Skerra, A., and Pluckthun, A. (1988) Assembly of a functional immunoglobulin Fv fragment in Escherichia coli. Science 240, 1038–1041.

    PubMed  CAS  Google Scholar 

  101. Fuchs, P., Dübel, S., Breitling, F., Braunagel, M., Klewinghaus, I., and Little, M. (1992) Recombinant human monoclonal antibodies. Basic principles of the immune system transferred to E. coli. Cell Biophys. 21, 81–91.

    PubMed  CAS  Google Scholar 

  102. Ward, E. S., Güssow, D., Griffiths, A. D., Jones, P. T., and Winter, G. (1989) Binding activities of a repertoire of single immunoglobulin variable domains secreted from E. coli. Nature 341, 544–546.

    PubMed  CAS  Google Scholar 

  103. Huse, W. D., Sastry, L., Iverson, S. A., Kang, A. S., Alting-Mees, M., Burton, D. R., Benkovic, S. J., and Lerner, R. A. (1989) Generation of a large combinatorial library of the immunoglobin repertoire in phage lambda. Science 246, 1275–1281.

    PubMed  CAS  Google Scholar 

  104. Caton, A. J., and Koprowski, H. (1990) Influenza virus hemagglutinin-specific antibodies isolated from a combinatorial expression library are closely related to the immune response of the donor. Proc. Natl. Acad. Sci. USA 87, 6450–6454.

    PubMed  CAS  Google Scholar 

  105. Mullinax, R. L., Gross, E. A., Amberg, J. R., Hay, B. N., Hogrefe, H. H., Kubitz, M. M., Greener, A., Alting-Mees, M., Ardourel, D., Short, J. M., Sorge, J. A., and Shopes, B. (1990) Identification of human antibody clones specific for tetanus toxoid in a bacteriophage lambda immunoexpression library. Proc. Natl. Acad. Sci. USA 87, 8095–8099.

    PubMed  CAS  Google Scholar 

  106. Russel, S. J., Hawkins, R. E., and Winter, G. (1993) Retroviral vectors displaying functional antibody fragments. Nucleic Acids Res. 21, 1081–1085.

    Google Scholar 

  107. Boublik, Y., Di Bonito, P., and Jones, I. M. (1995) Eukaryotic virus display: engineering the major surface glycoprotein of the Autographa californica nuclear polyhedrosis virus (AcNPV) for the presentation of foreign proteins on the virus surface. Bio/Technology 13, 1079–1084.

    PubMed  CAS  Google Scholar 

  108. Kieke, M. C., Cho, B. K., Boder, E. T., Kranz, D. M., and Wittrup, K. D. (1997) Isolation of anti-T cell receptor scFv mutants by yeast surface display. Protein Eng. 10, 1303–1310.

    PubMed  CAS  Google Scholar 

  109. Mattheakis, L. C., Bhatt, R. R., and Dower, W. J. (1994) An in vitro polysome display system for identifying ligands from very large peptide libraries. Proc. Natl. Acad. Sci. USA 91, 9022–9026.

    PubMed  CAS  Google Scholar 

  110. Hanes, J., and Plückthun, A. (1997) In vitro selection and evolution of functional proteins by using ribosome display. Proc. Natl. Acad. Sci. USA 94, 4937–4942.

    PubMed  CAS  Google Scholar 

  111. He, M., and Taussig, M. J. (1997) Antibody-ribosome-mRNA (ARM) complexes as efficient selection particles for in vitro display and evolution of antibody combining sites. Nucl. Acids Res. 25, 5132–5134.

    PubMed  CAS  Google Scholar 

  112. Smith, G. P., and Petrenko, V. A. (1997) Phage display. Chem. Rev. 97, 391–410.

    PubMed  CAS  Google Scholar 

  113. Webster, R. E., and Lopez, J. (1985) Structure and assembly of the class I filamentous bacteriophage, in Virus Structure and Assembly (Casjens, S., eds.), Jones and Bartlett Inc., Boston/Portala Valley, USA, pp. 235–267.

    Google Scholar 

  114. Smith, G. P. (1988) Filamentous phages as cloning vectors, in: Vectors (Rodriguez, R. L., and Denhardt, D. T., eds.), Butterworths, Boston, MA, pp. 61–83.

    Google Scholar 

  115. Riechmann, L., and Holliger, P. (1997) The C-terminal domain of TolA is the coreceptor for filamentous phage infection of E. coli. Cell 90, 351–360.

    PubMed  CAS  Google Scholar 

  116. Smith, G. P. (1985) Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 228, 1315–1317.

    PubMed  CAS  Google Scholar 

  117. Parmley, S. F., and Smith, G. P. (1988) Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. Gene 73, 305–318.

    PubMed  CAS  Google Scholar 

  118. Scott, J. K., and Smith, G. P. (1990) Searching for peptide ligands with an epitope library. Science 249, 386–390.

    PubMed  CAS  Google Scholar 

  119. Devlin, J. J., Panganiban, L. C., and Devlin, P. E. (1990) Random peptide libraries: a source of specific protein binding molecules. Science 249, 404–406.

    PubMed  CAS  Google Scholar 

  120. Cwirla, S. E., Peters, E. A., Barrett, R. W., and Dower, W. J. (1990) Peptides on phage: A vast library of peptides for identifying ligands. Proc. Natl. Acad. Sci. USA 87, 6378–6382.

    PubMed  CAS  Google Scholar 

  121. McCafferty, J., Griffiths, A. D., Winter, G., and Chiswell, D. J. (1990) Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348, 552–554.

    PubMed  CAS  Google Scholar 

  122. Hoogenboom, H. R., Griffiths, A. D., Johnson, K. S., Chiswell, D. J., Hudson, P., and Winter, G. (1991) Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains. Nucl. Acids Res. 19, 4133–4137.

    PubMed  CAS  Google Scholar 

  123. Breitling, F., Dübel, S., Seehaus, T., Klewinghaus, I., and Little, M. (1991) A surface expression vector for antibody screening. Gene 104, 147–153.

    PubMed  CAS  Google Scholar 

  124. Barbas, C. F., Kang, A. S., Lerner, R. A., and Benkovic, S. J. (1991) Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc. Natl. Acad. Sci. USA 88, 7978–7982.

    PubMed  CAS  Google Scholar 

  125. Ørum, H., Andersen, P. S., Øster, A., Johansen, L. K., Riise, E., Bjørnvad, M., Svendsen, I., and Engberg, J. (1993) Efficient method for constructing comprehensive murine Fab antibody libraries displayed on phage. Nucl. Acids Res. 21, 4491–4498.

    PubMed  Google Scholar 

  126. Ward, R. L., Clark, M. A., Lees, J., and Hawkins, N. J. (1996) Retrieval of human antibodies from phagedisplay libraries using enzymatic cleavage. J. Immunol. Methods 189, 73–82.

    Google Scholar 

  127. Meulemans, E. V., Slobbe, R., Wasterval, P., Ramaekers, F. C. S., and Van Eys, G. J. J. M. (1994) Selection of phage-displayed antibodies specific for a cytoskeletal antigen by competitive elution with a monoclonal antibody. J. Mol. Biol. 244, 353–360.

    PubMed  CAS  Google Scholar 

  128. Dübel, S., Breitling, F., Klewinghaus, I., and Little, M. (1992) Regulated secretion and purification of recombinant antibodies in E. coli. Cell Biophys. 21, 69–79.

    PubMed  Google Scholar 

  129. Hayashi, N., Kipriyanov, S., Fuchs, P., Welschof, M., Dörsam, H., and Little, M. (1995) A single expression system for the display, purification and conjugation of single-chain antibodies. Gene 160, 129–130.

    PubMed  CAS  Google Scholar 

  130. Waterhouse, P., Griffiths, A., Johnson, K. S., and Winter, G. (1993) Combinatorial infection and in vivo recombination: a strategy for making large phage antibody repertoires. Nucl. Acids Res. 21, 2265–2266.

    PubMed  CAS  Google Scholar 

  131. Geoffroy, F., Sodoyer, R., and Aujame, L. (1994) A new phage display system to construct multicombinatorial libraries of very large antibody repertoires. Gene 151, 109–113.

    PubMed  CAS  Google Scholar 

  132. Little, M., Fuchs, P., Breitling, F., and Dübel, S. (1993) Bacterial surface presentation of proteins and peptides: an alternative to phage technology? Trends Biotechnol. 11, 3–5.

    PubMed  CAS  Google Scholar 

  133. Duenas, M., and Borrebaeck, C. A. (1994) Clonal selection and amplification of phage displayed antibodies by linking antigen recognition and phage replication. Bio/Technology 12, 999–1002.

    PubMed  CAS  Google Scholar 

  134. Krebber, C., Spada, S., Desplancq, D., and Plückthun, A. (1995) Co-selection of cognate antibody-antigen pairs by selectively-infective phages. FEBS Lett. 377, 227–231.

    PubMed  CAS  Google Scholar 

  135. Krebber, C., Spada, S., Desplancq, D., Krebber, A., Ge, L., and Plückthun, A. (1997) Selectively-infective phage (SIP): a mechanistic dissection of a novel in vivo selection for protein-ligand interactions. J. Mol. Biol. 268, 607–618.

    PubMed  CAS  Google Scholar 

  136. Rondot, S., Anthony, K., Dübel, S., Ida, N., Beyreuther, K., Frost, L., Little, M., and Breitling, F. (1998) Epitopes fused to F-pilin are incorporated into functional recombinant pili. J. Mol. Biol. 279, 589–603.

    PubMed  CAS  Google Scholar 

  137. Malmborg, A. C., Söderlind, E., Frost, L., and Borrebaeck, C. A. (1997) Selective phage infection mediated by epitope expression on F pilus. J. Mol. Biol. 273, 544–551.

    PubMed  CAS  Google Scholar 

  138. Bass, S., Greene, R., and Wells, J. A. (1990) Hormone phage: an enrichment method for variant proteins with altered binding properties. Proteins 8, 309–314.

    PubMed  CAS  Google Scholar 

  139. Kuhn, A., and Wickner, W. (1985) Isolation of mutants in M13 coat protein that affect its synthesis, processing and assembly into phage. J. Biol. Chem. 260, 15,907–15,913.

    CAS  Google Scholar 

  140. Felici, F., Castagnoli, L., Musacchio, A., Jappelli, R., and Cesareni, G. (1991) Selection of antibody ligands from a large library of oligopeptides expressed on a multivalent exposition vector. J. Mol. Biol. 222, 301–310.

    PubMed  CAS  Google Scholar 

  141. Ilyichev, A. A., Minenkova, O. O., Kishchenko, G. P., Tat’kov, S. I., Karpishev, N. N., Eroshkin, A. M., Ofitzerov, V. I., Akimenko, Z. A., Petrenko, V. A., and Sandakhchiev, L. S. (1992) Inserting foreign peptides into the major coat protein of bacteriophage M13. FEBS Lett. 301, 322–324.

    PubMed  CAS  Google Scholar 

  142. Kang, A. S., Barbas, C. F., Janda, K. D., Benkovic, S. J., and Lerner, R. A. (1991) Linkage of recognition and replication functions by assembling combinatorial antibody Fab libraries along phage surfaces. Proc. Natl. Acad. Sci. USA 88, 4363–4366.

    PubMed  CAS  Google Scholar 

  143. Huse, W. D., Stinchcombe, T. J., Glaser, S. M., Starr, L., MacLean, M., Hellström, K. E., Hellström, I., and Yelton, D. E. (1992) Application of a filamentous phage pVIII fusion protein system suitable for efficient production, screening, and mutagenesis of F(ab’) antibody fragments. J. Immunol. 149, 3914–3920.

    PubMed  CAS  Google Scholar 

  144. Gram, H., Marconi, L. A., Barbas, C. F., Collet, T. A., Lerner, R. A., and Kang, A. S. (1992) In vitro selection and affinity maturation of antibodies from a naive combinatorial immunoglobulin library. Proc. Natl. Acad. Sci. USA 89, 3576–3580.

    PubMed  CAS  Google Scholar 

  145. Kretzschmar, T., and Geiser, M. (1995) Evaluation of antibodies fused to minor coat protein III and major coat protein VIII of bacteriophage M13. Gene 155, 61–65.

    PubMed  CAS  Google Scholar 

  146. Simons, G. F. M., Konings, R. N. H., and Schoenmakers, J. G. G. (1981) Genes VI, VII and IX of phage M13 code for minor capsid proteins of the virion. Proc. Natl. Acad. Sci. USA 78, 4194–4198.

    PubMed  CAS  Google Scholar 

  147. Makowski, L. (1992) Terminating a macromolecular helix: structural model for the minor proteins of bacteriophage M13. J. Mol. Biol. 228, 885–892.

    PubMed  CAS  Google Scholar 

  148. Jespers, L. S., Messens, J. H., De Keyser, A., Eeckhout, D., Van Den Brande, I., Gansemans, Y. G., Lauwereys, M. J., Vlasuk, G. P., and Stanssens, E. (1995) Surface expression and ligand-based selection of cDNAs fused to filamentous phage gene VI. Bio/Technology 13, 378–382.

    PubMed  CAS  Google Scholar 

  149. Hogrefe, H. H., Mullinax, R. L., Lovejoy, A. E., Hay, B. N., and Sorge, J. A. (1993) A bacteriophage lambda vector for the cloning and expression of immunoglobulin Fab fragments on the surface of filamentous phage. Gene 128, 119–126.

    PubMed  CAS  Google Scholar 

  150. Brinkmann, U., Chowdhury, P. S., Roscoe, D. M., and Pastan, I. (1995) Phage display of disulfide-stabilized Fv fragments. J. Immunol. Methods 182, 41–50.

    PubMed  CAS  Google Scholar 

  151. McGuinness, B. T., Walter, G., FitzGerald, K., Schuler, P., Mahoney, W., Duncan, A. R., and Hoogenboom, H. R. (1996) Phage diabody repertoires for selection of large numbers of bispecific antibody fragments. Nature Biotechnol. 14, 1149–1154.

    Google Scholar 

  152. Söderlind, E., Lagerkvist, A. C. S., Duenas, M., Malmborg, A.-C., Ayala, M., Danielsson, L., and Borrebaeck, C. A. K. (1993) Chaperonin assisted phage display of antibody fragments on filamentous bacteriophages. Bio/Technology 11, 503–507.

    PubMed  Google Scholar 

  153. Bothmann, H., and Plückthun, A. (1998) Selection for a periplasmic factor improving phage display and functional periplasmic expression. Nature Biotechnol. 16, 376–380.

    CAS  Google Scholar 

  154. Fuchs, P., Breitling, F., Dübel, S., Seehaus, T., and Little, M. (1991) Targeting recombinant antibodies to the surface of Escherichia coli: fusion to a peptidoglycan associated lipoprotein. Bio/Technology 9, 1369–1372.

    PubMed  CAS  Google Scholar 

  155. Fuchs, P., Weichel, W., Dübel, S., Breitling, F., and Little, M. (1996) Separation of E. coli expressing functional cell-wall bound antibody fragments by FACS. Immunotechnology 2, 97–102.

    Google Scholar 

  156. Francisco, J. A., Campbell, R., Iverson, B. L., and Georgiou, G. (1993) Production and fluorescence-activated cell sorting of Escherichia coli expressing a functional antibody fragment on the external surface. Proc. Natl. Acad. Sci. USA 90, 10,444–10,448.

    CAS  Google Scholar 

  157. Lu, Z., Murray, K. S., Van Cleave, V., LaVallie, E. R., Stahl, M., and McCoy, J. M. (1995) Expression of thioredoxin random peptide libraries on the Escherichia coli cell surface as functional fusions to flagellin: a system designed for exploring protein-protein interactions. Bio/Technology 13, 366–372.

    PubMed  CAS  Google Scholar 

  158. Bradbury, A., Persic, L., Werge, T., and Cattaneo, A. (1993) Use of living columns to select specific phage antibodies. Bio/Technology 11, 1565–1569.

    PubMed  CAS  Google Scholar 

  159. Davies, E. L., Smith, J. S., Birkett, C. R., Manser, J. M., Anderson-Dear, D. V., and Young, J. R. (1995) Selection of specific phage-display antibodies using libraries derived from chicken immunoglobulin genes. J. Immunol. Methods 186, 125–135.

    PubMed  CAS  Google Scholar 

  160. Arbabi Ghahroudi, M., Desmyter, A., Wyns, L., Hamers, R., and Muyldermans, S. (1997) Selection and identification of single domain antibody fragments from camel heavy-chain antibodies. FEBS Lett. 414, 521–526.

    PubMed  CAS  Google Scholar 

  161. Orlandi, R., Gussow, D. H., Jones, P. T., and Winter, G. (1989) Cloning immunoglobulin variable domains for expression by the polymerase chain reaction. Proc. Natl. Acad. Sci. USA 86, 3833–3837.

    PubMed  CAS  Google Scholar 

  162. Persson, M. A. A., Caothien, R. H., and Burton, D. R. (1991) Generation of diverse high-affinity human monoclonal antibodies by repertoire cloning. Proc. Natl. Acad. Sci. USA 88, 2432–2436.

    PubMed  CAS  Google Scholar 

  163. Chester, K. A., Begent, R. H. J., Robson, L., Keep, P., Pedley, R. B., Boden, J. A., Boxer, G., Green, A., Winter, G., Cochet, O., and Hawkins, R. E. (1994) Phage libraries for generation of clinically useful antibodies. Lancet 343, 455,456.

    PubMed  CAS  Google Scholar 

  164. Amas, R. S., Tornetta, M. A., Jones, C. S., and Tsui, P. (1994) Isolation of neutralizing anti-C5a monoclonal antibodies from a filamentous phage monovalent Fab display library. J. Immunol. 152, 4572–4581.

    Google Scholar 

  165. Burton, D. R., Barbas, C. F. d., Persson, M. A., Koenig, S., Chanock, R. M., and Lerner, R. A. (1991) A large array of human monoclonal antibodies to type 1 human immunodeficiency virus from combinatorial libraries of asymptomatic seropositive individuals. Proc. Natl. Acad. Sci. USA 88, 10,134–10,137.

    CAS  Google Scholar 

  166. Barbas, C. F., Bjorling, E., Chiodi, F., Dunlop, N., Cababa, D., Jones, T. M., Zebedee, S. L., Persson, M. A., Nara, P. L., Norrby, E., and Burton, D. R. (1992) Recombinant human Fab fragments neutralize human type 1 immunodeficiency virus in vitro. Proc. Natl. Acad. Sci. USA 89, 9339–9343.

    PubMed  CAS  Google Scholar 

  167. Barbas, C. F., Collet, T. A., Amberg, W., Roben, P., Binley, J. M., Hoekstra, D., Cababa, D., Jones, T. M., Williamson, R. A., Pilkington, G. R., Haigwood, N. L., Cabezas, E., Satterthwait, A. C., Sanz, I., and Burton, D. R. (1993) Molecular profile of an antibody response to HIV-1 as probed by combinatorial libraries. J. Mol. Biol. 230, 812–823.

    PubMed  CAS  Google Scholar 

  168. Barbas, C. F., Crowe, J. E., Cababa, D., Jones, T. M., Zebedee, S. L., Murphy, B. R., Chanock, R. M., and Burton, D. R. (1992) Human monoclonal Fab fragments derived from a combinatorial library bind to respiratory syncytial virus F glycoprotein and neutralize infectivity. Proc. Natl. Acad. Sci. USA 89, 10,164–10,168.

    CAS  Google Scholar 

  169. Zebedee, S. L., Barbas, C. F., Hom, Y. L., Caothien, R. H., Graff, R., DeGraw, J., Pyati, J., LaPolla, R., Burton, D. R., Lerner, R. A., and Thornton, G. B. (1992) Human combinatorial antibody libraries to hepatitis B surface antigen. Proc. Natl. Acad. Sci. USA 89, 3175–3179.

    PubMed  CAS  Google Scholar 

  170. Williamson, R. A., Burioni, R., Sanna, P. P., Partridge, L. J., Barbas, C. F., and Burton, D. R. (1993) Human monoclonal antibodies against a plethora of viral pathogens from single combinatorial libraries. Proc. Natl. Acad. Sci. USA 90, 4141–4145.

    PubMed  CAS  Google Scholar 

  171. Welschof, M., Terness, P., Kipriyanov, S. M., Stanescu, D., Breitling, F., Dörsam, H., Dübel, S., Little, M., and Opelz, G. (1997) The antigen-binding domain of a human IgG-anti-F(ab’)2 autoantibody. Proc. Natl. Acad. Sci. USA 94, 1902–1907.

    PubMed  CAS  Google Scholar 

  172. Ditzel, H. J., Binley, J. M., Moore, J. P., Sodroski, J., Sullivan, N., Sawyer, L. S. W., Hendry, R. M., Yang, W. P., Barbas, C. F., and Burton, D. R. (1995) Neurtralizing recombinant human antibodies to a conformational V2- and CD4-binding site-sensitive epitope of HIV-1 gp 120 isolated by using an epitope-masking procedure. J. Immunol. 154, 893–906.

    PubMed  CAS  Google Scholar 

  173. Little, M., Breitling, F., Dübel, S., Fuchs, P., Braunagel, M., Seehaus, T., and Klewinghaus, I. (1993) Universal antibody libraries on phage and bacteria, in Year Immunol., Vol. 7: Generation of antibodies by cell and gene immortalization (Terhorst, C., Malavasi, F., and Albertini, A., eds.), Karger, Basel, pp. 50–55.

    Google Scholar 

  174. Little, M., Breitling, F., Dubel, S., Fuchs, P., and Braunagel, M. (1995) Human antibody libraries in Escherichia coli. J. Biotechnol. 41, 187–195.

    PubMed  CAS  Google Scholar 

  175. Dörsam, H., Rohrbach, P., Kürschner, T., Kipriyanov, S., Renner, S., Braunagel, M., Welschof, M., and Little, M. (1997) Antibodies to steroids from a small human naive IgM library. FEBS Lett. 414, 7–13.

    PubMed  Google Scholar 

  176. Griffiths, A. D., Malmqvist, M., Marks, J. D., Bye, J. M., Embleton, M. J., McCafferty, J., Baier, M., Holliger, K. P., Gorick, B. D., Hughes-Jones, N. C., Hoogenboom, H. R., and Winter, G. (1993) Human anti-self antibodies with high specificity from phage display libraries. EMBO J. 12, 725–734.

    PubMed  CAS  Google Scholar 

  177. Schier, R., Marks, J. D., Wolf, E. J., Apell, G., Wong, C., McCartney, J. E., Bookman, M. A., Huston, J. S., Houston, L. L., Weiner, L. M., and Adams, G. P. (1995) In vitro and in vivo characterization of a human anti-c-erbB-2 single-chain Fv isolated from a filamentous phage antibody library. Immunotechnology 1, 73–81.

    PubMed  CAS  Google Scholar 

  178. Low, N. M., Holliger, P., and Winter, G. (1996) Mimicking somatic hypermutation: affinity maturation of antibodies displayed on bacteriophage using a bacterial mutator strain. J. Mol. Biol. 260, 359–368.

    Google Scholar 

  179. Hawkins, R. E., Russell, S. J., and G., W. (1992) Selection of phage antibodies by binding affinity. Mimicking affinity maturation. J. Mol. Biol. 226, 889–896.

    PubMed  CAS  Google Scholar 

  180. Riechmann, L., and Weill, M. (1993) Phage display and selection of a site-directed randomized single-chain antibody Fv fragment for its affinity improvement. Biochemistry 32, 8848–8855.

    PubMed  CAS  Google Scholar 

  181. Balint, R. F., and Larrick, J. W. (1993) Antibody engineering by parsimonious mutagenesis. Gene 137, 109–118.

    PubMed  CAS  Google Scholar 

  182. Barbas, C. F., Hu, D., Dunlop, N., Sawyer, L., Cababa, D., Hendry, R. M., Nara, P. L., and Burton, D. R. (1994) In vitro evolution of a neutralizing human antibody to human immunodeficiency virus type 1 to enhance affinity and broaden strain cross-reactivity. Proc. Natl. Acad. Sci. USA 91, 3809–3813.

    PubMed  CAS  Google Scholar 

  183. Yang, W.-P., Green, K., Pinz-Sweeney, S., Briones, A. T., Burton, D. R., and Barbas, C. F. (1995) CDR walking mutagenesis for the affinity maturation of a potent human anti-HIV-1 antibody into the picomolar range. J. Mol. Biol. 254, 392–403.

    PubMed  CAS  Google Scholar 

  184. Schier, R., McCall, A., Adams, G. P., Marshall, K. W., Merritt, H., Yim, M., Crawford, R. S., Weiner, L. M., Marks, C., and Marks, J. D. (1996) Isolation of picomolar affinity anti-c-erbB-2 single-chain Fv by molecular evolution of the complementarity determining regions in the center of the antibody binding site. J. Mol. Biol. 263, 551–567.

    Google Scholar 

  185. Marks, J. D., Hoogenboom, H. R., Griffiths, A. D., and Winter, G. (1992) Molecular evolution of proteins on filamentous phage. Mimicking the strategy of the immune system. J. Biol. Chem. 267, 16,007–16,010.

    CAS  Google Scholar 

  186. Schier, R., Bye, J., Apell, G., McCall, A., Adams, G. P., Malmqvist, M., Weiner, L. M., and Marks, J. D. (1996) Isolation of high-affinity monomeric human anti-c-erbB-2 single chain Fv using affinity-driven selection. J. Mol. Biol. 255, 28–43.

    Google Scholar 

  187. Thompson, J., Pope, T., Tung, J.-S., Chan, C., Hollis, G., Mark, G., and Johnson, K. S. (1996) Affinity maturation of a high-affinity human monoclonal antibody against the third hypervariable loop of human immunodeficiency virus: use of phage display to improve affinity and broaden strain reactivity. J. Mol. Biol. 256, 77–88.

    Google Scholar 

  188. Jespers, L. S., Roberts, A., Mahler, S. M., Winter, G., and Hoogenboom, H. R. (1994) Guiding the selection of human antibodies from phage display repertoires to a single epitope of an antigen. Biol Technology 12, 899–903.

    CAS  Google Scholar 

  189. Ohlin, M., Owman, H., Mach, M., and Borrebaeck, C. A. (1996) Light chain shuffling of a high affinity antibody results in a drift in epitope recognition. Mol. Immunol. 33, 47–56.

    Google Scholar 

  190. Kussie, P. H., Parhami-Seren, B., Wysocki, L. J., and Margolies, M. N. (1994) A single engineered amino acid substitution changes antibody fine specificity. J. Immunol. 152, 146–152.

    PubMed  CAS  Google Scholar 

  191. Ward, E. S. (1995) VH shuffling can be used to convert an Fv fragment of anti-hen lysozyme specificity to one that recognizes a T cell receptor Va. Mol. Immunol. 32, 147–156.

    PubMed  CAS  Google Scholar 

  192. Vaughan, T. J., Williams, A. J., Pritchard, K., Osbourn, J. K., Pope, A. R., Earnshaw, J. C., McCafferty, J., Hodits, R. A., Wilton, J., and Johnson, K. S. (1996) Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library. Nature Biotechnol. 14, 309–314.

    Google Scholar 

  193. Chothia, C., and Lesk, A. M. (1987) Canonical structures for the hypervariable regions of immunoglobulins. J.Mol.Biol. 196, 901–917.

    PubMed  CAS  Google Scholar 

  194. Chothia, C., Lesk, A. M., Tramontano, A., Levitt, M., Smith-Gill, S. J., Air, G., Sheriff, S., Padlan, E. A., Davies, D., Tulip, W. R., Colman, P. M., Spinelli, S., Alzari, P. M., and Poljak, R. J. (1989) Conformations of immunoglobulin hypervariable regions. Nature 342, 877–883.

    PubMed  CAS  Google Scholar 

  195. Chothia, C., Lesk, A. M., Gherardi, E., Tomlinson, I. M., Walter, G., Marks, J. D., Llewelyn, M. B., and Winter, G. (1992) Structural repertoire of the human VH segments. J. Mol. Biol. 227, 799–817.

    PubMed  CAS  Google Scholar 

  196. Vargas-Madrazo, E., Lara-Ochoa, F., and Almagro, J. C. (1995) Canonical structure repertoire of the antigen-binding site of immunoglobulins suggests strong geometrical restrictions associated to the mechanism of immune recognition. J. Mol. Biol. 254, 497–504.

    PubMed  CAS  Google Scholar 

  197. Barbas, C. F., Bain, J. D., Hoekstra, D. M., and Lerner, R. A. (1992) Semisynthetic combinatorial antibody libraries: a chemical solution to the diversity problem. Proc. Natl. Acad. Sci. USA 89, 4457–4461.

    PubMed  CAS  Google Scholar 

  198. Braunagel, M., and Little, M. (1997) Construction of a semisynthetic antibody library using trinucleotide oligos. Nucl. Acids Res. 25, 4690–4691.

    PubMed  CAS  Google Scholar 

  199. Barbas, C. F., Languino, L. R., and Smith, J. W. (1993) High-affinity self-reactive human antibodies by design and selection: targeting the integrin ligand binding site. Proc. Natl. Acad. Sci. USA 90, 10,003–10,007.

    CAS  Google Scholar 

  200. Barbas, C. F., Amberg, W., Simoncsits, A., Jones, T. M., and Lerner, R. A. (1993) Selection of human antihapten antibodies from semisynthetic libraries. Gene 137, 57–62.

    PubMed  CAS  Google Scholar 

  201. Barbas, C. F., Rosenblum, J. S., and Lerner, R. A. (1993) Direct selection of antibodies that coordinate metals from semisynthetic combinatorial libraries. Proc. Natl. Acad. Sci. USA 90, 6385–6389.

    PubMed  CAS  Google Scholar 

  202. Janda, K. D., Lo, C. L., Li, T., Barbas, C. F., Wirsching, P., and Lerner, R. A. (1994) Direct selection for a catalytic mechanism from combinatorial antibody libraries. Proc. Natl. Acad. Sci. USA 91, 2532–2536.

    PubMed  CAS  Google Scholar 

  203. Hoogenboom, H. R., and Winter, G. (1992) By-passing immunisation. Human antibodies from synthetic repertoires of germline VH gene segments rearranged in vitro. J. Mol. Biol. 227, 381–388.

    PubMed  CAS  Google Scholar 

  204. Akamatsu, Y., Cole, M. S., Tso, J. Y., and Tsurushita, N. (1993) Construction of a human Ig combinatorial library from genomic V segments and synthetic CDR3 fragments. J. Immunol. 151, 4651–4659.

    PubMed  CAS  Google Scholar 

  205. Nissim, A., Hoogenboom, H. R., Tomlinson, I. M., Flynn, G., Midgley, C., Lane, D., and Winter, G. (1994) Antibody fragments from a, single pot’ phage display library as immunochemical reagents. EMBO J. 13, 692–698.

    PubMed  CAS  Google Scholar 

  206. de Kruif, J., Terstappen, L., Boel, E., and Logtenberg, T. (1995) Rapid selection of cell subpopulation-specific human monoclonal antibodies from a synthetic phage antibody library. Proc. Natl. Acad. Sci. USA 92, 3938–4392.

    PubMed  Google Scholar 

  207. de Kruif, J., Boel, E., and Logtenberg, T. (1995) Selection and application of human single chain Fv antibody fragments from a semi-synthetic phage antibody display library with designed CDR3 regions. J. Mol. Biol. 248, 97–105.

    PubMed  CAS  Google Scholar 

  208. Griffiths, A. D., Williams, S. C., Hartley, O., Tomlinson, I. M., Waterhouse, P., Crosby, W. L., Kontermann, R. E., Jones, P. T., Low, N. M., Allison, T. J., Prospero, T. D., Hoogenboom, H. R., Nissim, A., Cox, J. P. L., Harrison, J. L., Zaccolo, M., Gherardi, E., and Winter, G. (1994) Isolation of high affinity human antibodies directly from large synthetic repertoires. EMBO J. 13, 3245–3260.

    PubMed  CAS  Google Scholar 

  209. Garrard, L. J., and Henner, D. J. (1993) Selection of an anti-IGF-1 Fab from a Fab phage library created by mutagenesis of multiple CDR loops. Gene 128, 103–109.

    PubMed  CAS  Google Scholar 

  210. Hayashi, N., Welschof, M., Zewe, M., Braunagel, M., Dübel, S., Breitling, F., and Little, M. (1994) Simultaneous mutagenesis of antibody CDR regions by overlap extension and PCR. Biotechniques 17, 310–315.

    PubMed  CAS  Google Scholar 

  211. Virnekäs, B., Ge, L., Plückthun, A., Schneider, K. C., Wellnhofer, G., and Moroney, S. E. (1994) Trinucleotide phosphoramidites: ideal reagents for the synthesis of mixed oligonucleotides for random mutagenesis. Nucl. Acids Res. 22, 5600–5607.

    PubMed  Google Scholar 

  212. Huston, J. S., Mudgett Hunter, M., Tai, M. S., McCartney, J., Warren, F., Haber, E., and Oppermann, H. (1991) Protein engineering of single-chain Fv analogs and fusion proteins. Methods Enzymol. 203, 46–88.

    Article  PubMed  CAS  Google Scholar 

  213. Buchner, J., and Rudolph, R. (1991) Renaturation, purification and characterization of recombinant Fabfragments produced in Escherichia coli. Bio/Technology 9, 157–162.

    PubMed  CAS  Google Scholar 

  214. Buchner, J., Pastan, I., and Brinkmann, U. (1992) A method for increasing the yield of properly folded recombinant fusion proteins: single-chain immunotoxins from renaturation of bacterial inclusion bodies. Anal. Biochem. 205, 263–270.

    PubMed  CAS  Google Scholar 

  215. Kurucz, I., Titus, J. A., Jost, C. R., and Segal, D. M. (1995) Correct disulfide pairing and efficient refolding of detergent-solubilized single-chain Fv proteins from bacterial inclusion bodies. Mol. Immunol. 32, 1443–1452.

    PubMed  CAS  Google Scholar 

  216. Kurucz, I., Titus, J. A., Jost, C. R., Jacobus, C. M., and Segal, D. M. (1995) Retargeting of CTL by an efficiently refolded bispecific single-chain Fv dimer produced in bacteria. J. Immunol. 154, 4576–4582.

    PubMed  CAS  Google Scholar 

  217. Skerra, A. (1993) Bacterial expression of immunoglobulin fragments. Curr. Opin. Immunol. 5, 256–262.

    PubMed  CAS  Google Scholar 

  218. Brinkmann, U., Buchner, J., and Pastan, I. (1992) Independent domain folding of Pseudomonas exotoxin and single-chain immunotoxins: influence of interdomain connections. Proc. Natl. Acad. Sci. USA 89, 3075–3079.

    PubMed  CAS  Google Scholar 

  219. Newton, D. L., Nicholls, P. J., Rybak, S. M., and Youle, R. J. (1994) Expression and characterization of recombinant human eosinophil-derived neurotoxin and eosinophil-derived neurotoxin-anti-transferrin receptor sFv. J. Biol. Chem. 269, 26,739–26,745.

    CAS  Google Scholar 

  220. Newton, D. L., Xue, Y., Olson, K. A., Fett, J. W., and Rybak, S. M. (1996) Angiogenin single-chain immunofusions: influence of peptide linkers and spacers between fusion protein domains. Biochemistry 35, 545–553.

    Google Scholar 

  221. Brinkmann, U., Reiter, Y., Jung, S. H., Lee, B., and Pastan, I. (1993) A recombinant immunotoxin containing a disulfide-stabilized Fv fragment. Proc. Natl. Acad. Sci. USA 90, 7538–7542.

    PubMed  CAS  Google Scholar 

  222. Reiter, Y., Brinkmann, U., Jung, S. H., Pastan, I., and Lee, B. (1995) Disulfide stabilization of antibody Fv: computer predictions and experimental evaluation. Protein Eng. 8, 1323–1331.

    PubMed  CAS  Google Scholar 

  223. Webber, K. O., Reiter, Y., Brinkmann, U., Kreitman, R., and Pastan, I. (1995) Preparation and characterization of a disulfide-stabilized Fv fragment of the anti-Tac antibody: comparison with its single-chain analog. Mol. Immunol. 32, 249–258.

    PubMed  CAS  Google Scholar 

  224. Pugsley, A. P. (1993) The complete general secretory pathway in gram-negative bacteria. Microbiol. Rev. 57, 50–108.

    PubMed  CAS  Google Scholar 

  225. Whitlow, M., and Filpula, D. (1991) Single-chain Fv proteins and their fusion proteins. Methods: A Companion to Meth. Enzymol. 2, 97–105.

    CAS  Google Scholar 

  226. Kipriyanov, S. M., Dübel, S., Breitling, F., Kontermann, R. E., and Little, M. (1994) Recombinant single-chain Fv fragments carrying C-terminal cysteine residues: production of bivalent and biotinylated miniantibodies. Mol. Immunol. 31, 1047–158.

    PubMed  CAS  Google Scholar 

  227. Hockney, R. C. (1994) Recent developments in heterologous protein production in Escherichia coli. Trends Biotechnol. 12, 456–463.

    PubMed  CAS  Google Scholar 

  228. Plückthun, A. (1994) Antibodies from Escherichia coli, in Handbook of Experimental Pharmacology, Vol. 113: The Pharmacology of Monoclonal Antibodies (Rosenberg, M., and Moore, G. P., eds.), Springer-Verlag, Berlin, Heidelberg, pp. 269–315.

    Google Scholar 

  229. Knappik, A., Krebber, C., and Plückthun, A. (1993) The effect of folding catalysts on the in vivo folding process of different antibody fragments expressed in Escherichia coli. Bio/Technology 11, 77–83.

    PubMed  CAS  Google Scholar 

  230. Duenas, M., Vazquez, J., Ayala, M., Soderlind, E., Ohlin, M., Perez, L., Borrebaeck, C. A., and Gavilondo, J. V. (1994) Intra-and extracellular expression of an scFv antibody fragment in E. coli: effect of bacterial strains and pathway engineering using GroES/L chaperonins. Biotechniques 16, 476–477.

    PubMed  CAS  Google Scholar 

  231. Knappik, A., and Plückthun, A. (1995) Engineered turns of a recombinant antibody improve its in vivo folding. Protein Eng. 8, 81–89.

    PubMed  CAS  Google Scholar 

  232. Dill, K. A. (1990) Dominant forces in protein folding. Biochemistry 29, 7133–7155.

    PubMed  CAS  Google Scholar 

  233. Bowden, G. A., and Georgiou, G. (1990) Folding and aggregation of beta-lactamase in the periplasmic space of Escherichia coli. J. Biol. Chem. 265, 16,760–16,766.

    CAS  Google Scholar 

  234. Kiefhaber, T., Rudolph, R., Kohler, H. H., and Buchner, J. (1991) Protein aggregation in vitro and in vivo: a quantitative model of the kinetic competition between folding and aggregation. Bio/Technology 9, 825–829.

    PubMed  CAS  Google Scholar 

  235. Kipriyanov, S. M., Moldenhauer, G., and Little, M. (1997) High level production of soluble single chain antibodies in small-scale Escherichia coli cultures. J. Immunol. Methods 200, 69–77.

    PubMed  CAS  Google Scholar 

  236. George, A. J., Titus, J. A., Jost, C. R., Kurucz, I., Perez, P., Andrew, S. M., Nicholls, P. J., Huston, J. S., and Segal, D. M. (1994) Redirection of T cellmediated cytotoxicity by a recombinant single-chain Fv molecule. J. Immunol. 152, 1802–1811.

    PubMed  CAS  Google Scholar 

  237. Kipriyanov, S. M., Dübel, S., Breitling, F., Kontermann, R. E., Heymann, S., and Little, M. (1995) Bacterial expression and refolding of singlechain Fv fragments with C-terminal cysteines. Cell Biophys. 26, 187–204.

    PubMed  CAS  Google Scholar 

  238. Skerra, A., and Pluckthun, A. (1991) Secretion and in vivo folding of the Fab fragment of the antibody McPC603 in Escherichia coli: influence of disulphides and cis-prolines. Protein Eng. 4, 971–979.

    PubMed  CAS  Google Scholar 

  239. Takkinen, K., Laukkanen, M.-L., Sizmann, D., Alfthan, K., Immonen, T., Vanne, L., Kaartinen, M., Knowles, J. K. C., and Teeri, T. T. (1991) An active single-chain antibody containing a cellulase linker domain is secreted by Escherichia coli. Protein Eng. 4, 837–841.

    PubMed  CAS  Google Scholar 

  240. Kipriyanov, S. M., Moldenhauer, G., Martin, A. C., Kupriyanova, O. A., and Little, M. (1997) Two amino acid mutations in an anti-human CD3 single chain Fv antibody fragment that affect the yield on bacterial secretion but not the affinity. Protein Eng. 10, 445–453.

    PubMed  CAS  Google Scholar 

  241. Sawyer, J. R., Schlom, J., and Kashmiri, S. V. S. (1994) The effect of induction conditions on production of a soluble anti-tumor sFv in Escherichia coli. Protein Eng. 7, 1401–1406.

    PubMed  CAS  Google Scholar 

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  1. Recombinant Antibody Research Group (D0500), German Cancer Research Center, Im Neuenheimer Feld 280, D-69120, Heidelberg, Germany

    Melvyn Little

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Kipriyanov, S.M., Little, M. Generation of recombinant antibodies. Mol Biotechnol 12, 173–201 (1999). https://doi.org/10.1385/MB:12:2:173

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