Expression and Secretion of Repertoires of VH Domains in Escherichia Coli: Isolation of Antigen Binding Activities

Conference paper


The production of monoclonal antibodies using hybridoma technology (Köhler and Milstein, 1975) represents a landmark for the use of antibodies as reagents for medicine and biology. It is now possible to use recombinant DNA methods to isolate and genetically manipulate antibody genes, and the resulting immunoglobulins or immunoglobulin fragments can be efficiently expressed in mammalian or bacterial hosts (for a review, see Morrison et al., 1988).The immunoglobulin molecule consists of a string of domains, each domain consisting of about 100 amino acid residues. The constant domains carry the effector functions, such as complement mediated lysis and ADCC. The antigen binding site is fashioned by both heavy (VH) and light (VL; Vκ or Vλ) chain variable domains, as demonstrated by the solved crystallographic structures of antibody in association with antigen (Amit et al., 1986; Sheriff et al., 1987; Colman et al.,1987) or hapten (Satow et al, 1986). Variable domains have been pasted onto constant domains (Morrison et ah, 1984; Boulianne et al., 1984; Neuberger et al.,1985) and hypervariable loops (CDRs) onto the underlying (β-sheet framework of variable domains (Jones et al.,1986; Verhoeyen et al.,1988; Riechmann et al.,1988a). Grafting CDRs has been used to humanise rodent antibodies and one such antibody used in the successful treatment of 2 patients with non- Hodgkins lymphoma (Hale et al.,1988; Riechmann et al.,1988a).


Complement Mediate Lysis Keyhole Limpet Haemocyanin Antigen Binding Activity Hypervariable Loop Chain Variable Domain 
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  1. Amit AG, Mariuzza RA, Phillips SEV, Poljak RJ (1986) Science 233:747–754PubMedCrossRefGoogle Scholar
  2. Better M, Chang CP, Robinson RR, Horwitz AH (1988) Science 240:1041–1043PubMedCrossRefGoogle Scholar
  3. Boulianne GL, Hozumi N, Shulman MJ (1984) Nature 312:643–646PubMedCrossRefGoogle Scholar
  4. Chothia C, Lesk AM, Dodson GG, Hodgkin DC (1983) Nature 302:500–505PubMedCrossRefGoogle Scholar
  5. Colman PM, Laver WG, Varghese JN, Baker AT, Tulloch PA, Air GM, Webster RG (1987) Nature 326:358–362PubMedCrossRefGoogle Scholar
  6. Edmundson AB, Ely KR, Herron JN Molecular Immunology (1984) 21:561–576CrossRefGoogle Scholar
  7. Evan GI, Lewis GK, Ramsay G, Bishop JM (1985) Mol Cell Biol 5:3610–3616.PubMedCentralPubMedGoogle Scholar
  8. Fearnley IM, Runswick MJ, Walker, JE EMBO J (1989) 8:665–672PubMedCentralPubMedGoogle Scholar
  9. Fleischman JB, Porter RR, Press EM, Biochem J (1963) 88:220–228PubMedCentralPubMedGoogle Scholar
  10. Getzoff ED, Geysen HM, Rodda SJ, Alexander H, Tainer JA, Lerner RA Science (1987) 235:1191–1196Google Scholar
  11. Gronenborn B Molec Gen Gen (1976) 148:243–250CrossRefGoogle Scholar
  12. Hale G, Dyer MJS, Clark MR, Phillips JM, Marcus R, Riechmann L, Winter G, Waldmann H The Lancet (1988) December 17:1394–1399CrossRefGoogle Scholar
  13. Jaton J-C, Klinman NR, Givol D, Sela M Biochemistry (1968) 7:4185–4195Google Scholar
  14. Jones PT, Dear PH, Foote J, Neuberger MS, Winter G Nature (1986) 321:522–525PubMedCrossRefGoogle Scholar
  15. Kabat EA, Wu TT, Reid-Miller M, Gottesman KS in Sequences of Proteins of Immunological Interest (US Dept of Health and Human Services, US Government Printing Office, 1987) Köhler G, Milstein C Nature (1975) 256:495–497Google Scholar
  16. Laemmli UK Nature (London) (1970) 227:680–685CrossRefGoogle Scholar
  17. Levison SA, Kierszenbaum F, Dandliker WB Biochemistry (1970) 9:322–331PubMedCrossRefGoogle Scholar
  18. Maniatis T, Fritsch EF, Sambrook J (eds.) in “Molecular Cloning; a Laboratory Manual” (Cold Spring Harbour 1982) Matsudaira PJ Biol Chem (1987) 262:10035–10038Google Scholar
  19. Miller JH Experiments in Molecular Genetics, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York (1972)Google Scholar
  20. Morrison SL, Johnson MJ, Herzenberg LA, Oi VT Proc Nad Acad Sci USA (1984) 81:6351–6855Google Scholar
  21. Morrsion SL, Wims, LA, Wallick S, Tan L, Oi VT Ann NY Acad Sci (1988) 507:187–198CrossRefGoogle Scholar
  22. Munro S, Pelham H Cell (1986) 46:291–300PubMedCrossRefGoogle Scholar
  23. Neuberger MS, Williams GT, Mitchell EB, Jouhal SS, Flanagan JG, Rabbitts TH Nature (1985) 314:268–270PubMedCrossRefGoogle Scholar
  24. Orlandi R, Gussow DH, Jones PT, Winter G Proc Natl Acad Sci USA (1989) 86:3833–3837PubMedCentralPubMedCrossRefGoogle Scholar
  25. Riechmann L, Clark M, Waldmann H, Winter G Nature (1988a) 332:323–327PubMedCrossRefGoogle Scholar
  26. Riechmann L, Foote J, Winter G J Mol Biol (1988b) 203:825–828PubMedCrossRefGoogle Scholar
  27. Rossman MG et al., Nature (1985) 317:145–153.CrossRefGoogle Scholar
  28. Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N Science (1985) 230:1350–1354PubMedCrossRefGoogle Scholar
  29. Satow Y, Cohen GH, Padlan EA, Davies DR J Mol Biol (1986) 190:593–604.PubMedCrossRefGoogle Scholar
  30. Sheriff S, Silverton EW, Padlan EA, Cohen GH, Smith-Gill SJ, Finzel BC, Davies DR Proc Natl Acad Sci. USA (1987) 84:8075–8079PubMedCentralPubMedCrossRefGoogle Scholar
  31. Skerra A, Pluckthun A Science (1988)240:1038–1040PubMedCrossRefGoogle Scholar
  32. Towbin H, Staehelin T, Gordon J Proc Natl Acad Sci USA (1979) 76:4350–4354.PubMedCentralPubMedCrossRefGoogle Scholar
  33. Utsumi S, Karush F, Biochemistry (1964)3:1329–1338PubMedCrossRefGoogle Scholar
  34. Ward ES, Gussow DH, Griffiths A, Jones PT, Winter G (1989) submitted.Google Scholar
  35. Verhoeyen M, Milstein C, Winter G Science (1988) 239:1534–1536PubMedCrossRefGoogle Scholar
  36. Yanisch-Perron C, Vieira J, Messing J Gene (1985) 33:103–119.Google Scholar

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© Springer-Verlag Berlin Heidelberg 1989

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