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The production and application of single-chain antibody fragments

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Abstract

This review discusses methods for the single-chain antibody fragment (scFv) generation and scFv expression systems, and describes potential applications of scFv in the therapy of viral diseases and cancer, with emphasis on intracellularly expressed scFvs (intrabodies), application of scFvs in detection and diagnostics, and their use in proteomics.

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Abbreviations

CH :

heavy-chain constant domain

CL :

light-chain constant domain

ELISA:

enzyme-linked immunosorbent assay

ER:

endoplasmic reticulum

Fab:

monovalent antigen binding fragment

Fv:

variable-region fragment

HCV:

hepatitis C virus

IMAC:

immobilized metal affinity chromatography

LTR:

long terminal repeats

mAb:

monoclonal antibody

RRE:

rev responsible element

scFv:

single-chain antibody fragment

TAR:

transactivation response element

VH :

heavy-chain variable domain

VL :

light-chain variable domain

References

  • Alvarez R.D., Barnes M.N., Gomez-Navarro J., Wang M., Strong T.V., Arafat W., Arani R.B., Johnson M.R., Roberts B.L., Siegal G.P., Curiel D.T.: A cancer gene therapy approach utilizing an anti-erbB-2 single-chain antibody-encoding adenovirus (AD21): a phase I trial.Clin.Cancer Res. 6, 3081–3087 (2000).

    PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Bai J., Sui J., Zhu R.Y., Tallarico A.S., Gennari F., Zhang D., Marasco W.A.: Inhibition of Tat-mediated transactivation and HIV-1 replication by human anti-hCyclinT1 intrabodies.J.Biol.Chem. 278, 1433–1442 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Baltimore D.: Gene therapy. Interacellular immunization.Nature 335, 395–396 (1988).

    Article  PubMed  CAS  Google Scholar 

  • Barbas C.F. 3rd,Kang S., Lerner R.A., Benkovic S.J.: Assembly of combinatorial antibody libraries on phage surfaces: the gene III site.Proc.Nat.Acad.Sci.USA 88, 7978–7982 (1991).

    Article  PubMed  CAS  Google Scholar 

  • Barbas C.F. 3rd,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., Burton D.R.: Molecular profile of an antibody response to HIV-1 as probed by combinatorial libraries.J.Mol.Biol. 230, 812–823 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Bei R., Schlom J., Kashmiri S.V.: Baculovirus expression of a functional single-chain immunglobulin and its IL-2 fusion protein.J.Immunol.Meth. 186, 245–255 (1995).

    Article  CAS  Google Scholar 

  • Biocca S., Pierandrei-Amaldi P., Campioni N., Cattaneo A.: Intracellular immunization with cytosolic recombinant antibodies.Biotechnology (New York) 12, 396–399 (1994).

    Article  CAS  Google Scholar 

  • 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., Whitlow M.: Single-chain antigen-binding proteins.Science 242, 423–426 (1988).

    Article  PubMed  CAS  Google Scholar 

  • BouHamdan M., Duan L.X., Pomerantz R.J., Strayer D.S.: Inhibition of HIV-1 by an anti-integrase single-chain variable fragment (SFv): delivery by SV40 provides durable protection against HIV-1 and does not require selection.Gene Ther. 6, 660–666 (1999).

    Article  PubMed  CAS  Google Scholar 

  • BouHamdan M., Kulkosky J., Duan L.X., Pomerantz R.J.: Inhibition of HIV-1 replication and infectivity by expression of a fusion protein, VPR-anti-integrase single-chain variable fragment (SFv): intravirion molecular therapies.J.Human Virol. 3, 6–15 (2000).

    CAS  Google Scholar 

  • BouHamdan M., Strayer D.S., Wei D., Mukhtar M., Duan L.X., Hoxie J., Pomerantz R.J.: Inhibition of HIV-1 infection by down-regulation of the CXCR4 co-receptor using an intracellular single chain variable fragment against CXCR4.Gene Ther. 8, 408–418 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Brown P.O., Botstein D.: Exploring the new world of the genome with DNA microarrays.Nature Genet. 21, 33–37 (1999).

    Article  PubMed  CAS  Google Scholar 

  • Celer V., Blažek D., Navrátilová I., Skládal P., Blacklaws B., Bujdoso R.: Recombinant single-chain Fv antibodies that recognize the p25 protein of the Maedi-Visna virus.Folia Microbiol. 48, 435–440 (2003).

    Article  CAS  Google Scholar 

  • Chari R.V.: Targeted delivery of chemotherapeutics: tumor-activated prodrug therapy.Adv.Drug Deliv.Rev. 31, 89–104 (1998).

    Article  PubMed  CAS  Google Scholar 

  • Charlton K., Harris W.J., Porter A.J.: The isolation of super-sensitive anti-hapten antibodies from combinatorial antibody libraries derived from sheep.Biosens.Bioelectron. 16, 639–646 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Chen S.Y., Khouri Y., Bagley J., Marasco W.A.: Combined intra- and extracellular immunization against human immunodeficiency virus type 1 infection with a human anti-gp120 antibody.Proc.Nat.Acad.Sci.USA 91, 5932–5936 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Clackson T., Hoogenboom H.R., Griffiths A.D., Winter G.: Making antibody fragments using phase display libraries.Nature 352, 624–628 (1991).

    Article  PubMed  CAS  Google Scholar 

  • Cochet O., Gruel N., Fridman W.H., Teillaud J.L.: Ras and p53 intracellular targeting with recombinant single-chain Fv (scFv) fragments: a novel approach for cancer therapy?Cancer Detect.Prev. 23, 506–510 (1999).

    Article  PubMed  CAS  Google Scholar 

  • Conrad U., Fielder U.: Expression of engineered antibodies in plant cells.Plant Mol.Biol. 26, 1023–1030 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Davis G.T., Bedzyk W.D., Voss E.W., Jacobs T.W.: Single chain antibody (SCA) encoding genes: one-step construction and expression in eukaryotic cells.Biotechnology (New York) 9, 165–169 (1991).

    Article  CAS  Google Scholar 

  • Duan L., Bagasra O., Laughlin M.A., Oakes J.W., Pomerantz R.J.: Potent inhibition of human immunodeficiency virus type 1 replication by an intracellular anti-Rev single-chain antibody.Proc.Nat.Acad.Sci.USA 91, 5075–5079 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Freyre F.M., Vazquez J.E., Ayala M., Canaan-Haden L., Bell H., Rodriguez I., Gonzalez A., Cintado A., Gavilondo J.V.: Very high expression of an anti-carcinoembryonic antigen single chain Fv antibody fragment in the yeastPichia pastoris.J.Biotechnol. 76, 157–163 (2000).

    Article  PubMed  CAS  Google Scholar 

  • George A.J., Jamar F., Tai M.S., Heelan B.T., Adams G.P., McCartney J.E., Houston L.L., Weiner L.M., Oppermann H., Peters A.M.: Radiometal labeling of recombinant proteins by a genetically engineered minimal chelation site: technetium-99m coordination by single-chain Fv antibody fusion proteins through a C-terminal cysteinyl peptide.Proc.Nat.Acad.Sci.USA 92, 8358–8362 (1995).

    Article  PubMed  CAS  Google Scholar 

  • Goncalves J., Silva F., Freitas-Vieira A., Santa-Marta M., Malho R., Yang X., Gabuzda D., Barbas C., 3rd: Functional neutralization of HIV-1 Vif protein by intracellular immunization inhibits reverse transcription and viral replication.J.Biol.Chem. 277, 32036–32045 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Griep R.A., van Twisk C., Kerschbaumer R.J., Harper K., Torrance L., Himmler G., van der Wolf J.M., Schots A.: pSKAP/S: an expression vector for the production of single-chain Fv alkaline phosphatase fusion proteins.Prot.Exp.Purif. 16, 63–69 (1999a).

    Article  CAS  Google Scholar 

  • Griep R.A., van Twisk C., van der Wolf J.M., Schots A.: Fluobodies: green fluorescent single-chain Fv fusion proteins.J.Immunol.Meth. 230, 121–130 (1999b).

    Article  CAS  Google Scholar 

  • Griep R.A., Prins M., van Twisk C., Keller H.J.H.G., Kerschbaumer R.J., Kormelink R., Goldbach R.W., Schots A.: Application of phage display in selecting tomato spotted wilt virus-specific single-chain antibodies (scFvs) for sensitive diagnosis in ELISA.Phytopathology 90, 183–190 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Griffiths A.D., Duncan A.R.: Strategies for selection of antibodies by phage display.Curr.Opin.Biotechnol. 9, 102–108 (1998).

    Article  PubMed  CAS  Google Scholar 

  • Guirakhoo F., Catalan J., Monath T., Weltzin R.: Cloning, expression and functional activities of a single chain antibody fragment directed to fusion protein of respiratory syncytial virus.Immunotechnology 2, 219–228 (1996).

    Article  PubMed  CAS  Google Scholar 

  • de Haard H.J., Kazemier B., Koolen M.J., Nijholt L.J., Meloen R.H., van Gemen B., Hoogenboom H.R., Arends J.W.: Selection of recombinant, library-derived antibody fragments against p24 for human immunodeficiency virus type 1 diagnostics.Clin.Diagn.Lab.Immunol. 5, 636–644 (1998).

    PubMed  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Harper K., Toth R.L., Mayo M.A., Torrance L.: Properties of a panel of single chain variable fragments against potato leafroll virus obtained from two phage display libraries.J.Virol.Meth. 81, 159–168 (1999).

    Article  CAS  Google Scholar 

  • Heintges T., Zu Putlitz J., Wands J.R.: Characterization and binding of intracellular antibody fragments to the hepatitis C virus core protein.Biochem.Biophys.Res.Commun. 263, 410–418 (1999).

    Article  PubMed  CAS  Google Scholar 

  • Holt L.J., Bussow K., Walter G., Tomlinson I.M.: By-passing selection: direct screening for antibody-antigen interactions using protein arrays.Nucl.Acids Res. 28, E72 (2000).

    Google Scholar 

  • Hudson P.J., Souriau C.: Recombinant antibodies for cancer diagnosis and therapy.Expert.Opin.Biol.Ther. 1, 845–855 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Hust M., Maiss E., Jacobsen H.J., Reinard T.: The production of a genus-specific recombinant antibody (scFv) using a recombinant potyvirus protease.J.Virol.Meth. 106, 225–233 (2002).

    Article  CAS  Google Scholar 

  • Jost C.R., Kurucz I., Jacobus C.M., Titus J.A., George A.J., Segal D.M.: Mammalian expression and secretion of functional single-chain Fv molecules.J.Biol.Chem. 269, 26267–26273 (1994).

    PubMed  CAS  Google Scholar 

  • Jurado P., Ritz D., Beckwith J., de Lorenzo V., Fernandez L.A.: Production of functional single-chain Fv antibodies in the cytoplasm ofEscherichia coli.J.Mol.Biol. 320, 1–10 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Kalderon D., Richardson W.D., Markham A.F., Smith A.E.: Sequence requirements for nuclear location of simian virus 40 large-T antigen.Nature 311, 33–38 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Kerschbaumer R.J., Hirschl S., Kaufmann A., Ibl M., Koenig R., Himmler G.: Single-chain Fv fusion proteins suitable as coating and detecting reagents in a double antibody sandwich enzyme-linked immunosorbent assay.Anal.Biochem. 249, 219–227 (1997).

    Article  PubMed  CAS  Google Scholar 

  • Kipriyanov S.M., Moldenhauer G., Little M.: High level production of soluble single chain antibodies in small-scaleEscherichia coli cultures.J.Immunol.Meth. 200, 69–77 (1997).

    Article  CAS  Google Scholar 

  • Knappik A., Ge L., Honegger A., Pack P., Fischer M., Wellinhofer G., Hoess A., Wolle J., Pluckthun A., Virnekas B.: Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides.J.Mol.Biol. 296, 57–86 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Kohler G., Milstein C.: Continuous cultures of fused cell secreting antibody of predefined specificity.Nature 256, 495–497 (1975).

    Article  PubMed  CAS  Google Scholar 

  • Kozak M.: At least six nucleotides preceding the AUG initiator codon enhance translation in mammalian cells.J.Mol.Biol. 196, 947–950 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Kuroki M., Arakawa F., Khare P.D., Kuroki M., Liao S., Matsumoto H., Abe H., Imakhre T.: Specific targeting strategies of cancer gene therapy using a single-chain variable fragment (scFv) with a high affinity for CEA.Anticancer Res. 20, 4067–4071 (2000).

    PubMed  CAS  Google Scholar 

  • Legastelois I., Desgranges C.: Design and intracellular activity of a human single-chain antibody to human immunodeficiency virus type 1 conserved gp41 epitope.J.Virol. 74, 5712–5715 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Lener M., Horn I.R., Cardinale A., Messina S., Nielsen U.B., Rybak S.M., Hoogenboom H.R., Cattaneo A., Biocca S.: Diverting a protein from its cellular location by intracellular antibodies. The case of p21Ras.Eur.J.Biochem. 267, 1196–1205 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Levin R., Mhashilkar A.M., Dorfman T., Bukovsky A., Zani C., Bagley J., Hinkula J., Niedrig M., Albert J., Wahren B., Gottlinger H.G., Marasco W.A.: 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 (1997).

    PubMed  CAS  Google Scholar 

  • Levy-Mintz P., Duan L., Zhang H., Hu B., Dornadula G., Zhu M., Kulkosky J., Bizub-Bender D., Skalka A.M., Pomerantz R.J.: Intracellular expression of single-chain variable fragments to inhibit early stages of the viral life cycle by targeting human immunodeficiency virus type 1 integrase.J.Virol. 70, 8821–8832 (1996).

    PubMed  CAS  Google Scholar 

  • Li X., Stuckert P., Bosch I., Marks J.D., Marasco W.A.: Single-chain antibody-mediated gene delivery into ErbB2-positive human breast cancer cells.Cancer Gene Ther. 8, 555–565 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Li Y., Collins M.S., Whitelam G.C., Alexander D.J.: Rapid pathotyping of Newcastle disease virus using a single-chain Fv displayed on phage against the C-terminal end of the F2 polypeptide.Arch.Virol. 147, 2025–2037 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Liu B., Huang L., Sihlbom C., Burlingame A., Marks J.D.: Towards proteome-wide production of monoclonal antibody by phage display.J.Mol.Biol. 315, 1063–1073 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Liu S.Y., Fary J.F., Petersdorf S.H., Martin P.J., Maloney D.G., Appelbaum F.R., Matthews D.C., Bush S.A., Durack L.D., Fisher D.R., Gooley T.A., Bernstein I.D., Press O.W.: Follow-up of relapsed B-cell lymphoma patients treated with iodine-131-labeled anti-CD20 antibody and autologous stem-cell rescue.J.Clin.Oncol. 16, 3270–3278 (1998).

    PubMed  CAS  Google Scholar 

  • Marasco W.A.: Intrabodies: turning the humoral immune system outside in for intracellular immunization.Gene Ther. 4, 11–15 (1997).

    Article  PubMed  CAS  Google Scholar 

  • Marasco W.A., Dana Jones S.: Antibodies for targeted gene therapy: extracellular gene targeting and intracellular expression.Adv. Drug Deliv.Rev. 31, 153–170 (1998).

    Article  PubMed  Google Scholar 

  • Marasco W.A., Haseltine W.A., Chen S.Y.: Design intracellular expression, and activity of a human anti-human immunodeficiency virus type 1 gp120 single-chain antibody.Proc.Nat.Acad.Sci.USA 90, 7889–7893 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Marasco W.A., LaVecchio J., Winkler A.: Human anti-HIV-1 tat sFv intrabodies for gene therapy of advanced HIV-1-infection and AIDS.J.Immunol.Meth. 231, 223–238 (1999).

    Article  CAS  Google Scholar 

  • Marin M., Pelegrin-Zurilla M., Bachrach E., Noel D., Brockly F., Piechaczyk M.: Antiviral activity of an intracellularly expressed single-chain antibody fragment directed against the murine leukemia virus capsid protein.Human Gene Ther. 11, 389–401 (2000).

    Article  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  • Moradpour D., Bieck E., Hugle T., Wels W., Wu J.Z., Hong Z., Blum H.E., Bartenschlager R.: Functional properties of a monoclonal antibody inhibiting the hepatitis C virus RNA-dependent RNA polmerase.J.Biol.Chem. 277, 593–601 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Morino K., Katsumi H., Akahori Y., Iba Y., Shinohara M., Ukai Y., Kohara Y., Kurosawa Y.: Antibody fusions with fluorescent proteins: a versatile reagent for profiling protein expression.J.Immunol.Meth. 257, 175–184 (2001).

    Article  CAS  Google Scholar 

  • Muller B.H., Lafay F., Demangel C., Perrin P., Tordo N., Flamand A., Lafaye P., Guesdon J.L.: Phage-displayed and soluble mouse scFv fragments neutralize rabies virus.J.Virol.Meth. 67, 221–233 (1997).

    Article  CAS  Google Scholar 

  • Munro S., Pelham H.R.: A C-terminal signal prevents secretion of luminal ER proteins.Cell 48, 899–907 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Nejatollahi F., Hodgetts S.J., Vallely P.J., Burnie J.P.: Neutralizing human recombinant antibodies to human cytomegalovirus glycoproteins gB and gH.FEMS Immunol.Med.Microbiol. 34, 237–244 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Oelschlaeger P., Srikant-Iyer S., Lange S., Schmitt J., Schmid R.: Fluorophor-linked immunosorbent assay: a time- and cost-saving method for the characterization of antibody fragments using a fusion protein of a single-chain antibody fragment and enhanced green fluorescent protein.Anal.Biochem. 309, 27–34 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Owen M., Gandecha A., Cockburn B., Whitelam G.: Synthesis of a functional antiphytochrome single-chain-Fv protein in transgenic tobacco.Biotechnology 10, 790–794 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Pai L.H., Pastan I.: Clinical trials withPseudomonas exotoxin immunotoxins.Curr.Top.Microbiol.Immunol. 234, 83–96 (1998).

    PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Pluckthun A., Skerra A.: Expression of functional antibody Fv and Fab fragments inEscherichia coli.Meth.Enzymol. 178, 497–515 (1989).

    Article  PubMed  CAS  Google Scholar 

  • Ridder R., Geisse S., Kleuser B., Kawalleck P., Gram H.: A COS-cell-based system for rapid production and quantification of scFv::IgC-κ antibody fragments.Gene 166, 273–276 (1995).

    Article  PubMed  CAS  Google Scholar 

  • Rondon I.J., Marasco W.A.: Intracellular antibodies (intrabodies) for gene therapy of infectious diseases.Ann.Rev.Microbiol. 51, 257–283 (1997).

    Article  CAS  Google Scholar 

  • Shaheen F., Duan L., Zhu M., Bagasra O., Pomerantz R.J.: Targeting human immunodeficiency virus type 1 reverse transcriptase by intracellular expression of single-chain variable fragments to inhibit early stages of the viral life cycle.J.Virol. 70, 3392–3400 (1996).

    PubMed  CAS  Google Scholar 

  • Sheets M.D., Amersdorfer P., Finnern R., Sargent P., Lindquist E., Schier R., Hemingsen G., Wong C., Gerhart J.C., Marks J.D., Lindqvist E.: Efficient construction of a large nonimmune phage antibody library: the production of high-affinity human single-chain antibodies to protein antigens.Proc.Nat.Acad.Sci. USA 95, 6157–6162 (1998).

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Skerra A., Pluckthun A.: Assembly of a functional immunoglobulin Fv fragment inEscherichia coli.Science 240, 1038–1041 (1988).

    Article  PubMed  CAS  Google Scholar 

  • Steinberger P., Andris-Widhopf J., Buhler B., Torbett B.E., Barbas C.F. 3rd: Functional deletion of the CCR5 receptor by intracellular immunization produces cells that are refractory to CCR5-dependent HIV-1 infection and cell fusion.Proc.Nat.Acad.Sci.USA 97, 805–810 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Strayer D.S., Branco F., Landre J., BouHamdan M., Shaheen F., Pomerantz R.J.: Combination genetic therapy to inhibit HIV-1.Mol.Ther. 5, 33–41 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Strube R.W., Chen S.Y.: Characterization of anti-cyclin E single-chain Fv antibodies and intrabodies in breast cancer cells: enhanced intracellular stability of novel sFv-v-F(c) intrabodies.J.Immunol.Meth. 263, 149–167 (2002).

    Article  CAS  Google Scholar 

  • Sullivan D.E., Mondelli M.U., Curiel D.T., Krasnykh V., Mikheeva G., Gaglio P., Morris C.B., Dash S., Gerber M.A.: Construction and characterization of an intracellular single-chain human antibody to hepatitis C virus non-structural 3 protein.J.Hepatol. 37, 660–668 (2002).

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Thor Straten P., Guldberg P., Schrama D., Andersen M.H., Moerch U., Seremet T., Siedel C., Reisfeld R.A., Becker J.C.:In situ cytokine therapy: redistribution of clonally expanded T cells.Eur.J.Immunol. 31, 250–258 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Verma R., Boleti E., George A.J.: Antibody engineering: comparison of bacterial, yeast, insect and mammalian expression systems.J.Immunol.Meth. 216, 165–181 (1998).

    Article  CAS  Google Scholar 

  • de Wildt R.M., Mundy C.R., Gorick B.D., Tomlinson I.M.: Antibody arrays for high-throughput screening of antibody-antigen interactions.Nat.Biotechnol. 18, 989–994 (2000).

    Article  PubMed  Google Scholar 

  • Wu Y., Duan L., Zhu M., Hu B., Kubota S., Bagasra O., Pomerantz R.J.: Binding of intracellular anti-Rev single chain variable fragments to different epitopes of human immunodeficiency virus type 1 rev: variations in viral inhibition.J.Virol. 70, 3290–3297 (1996).

    PubMed  CAS  Google Scholar 

  • Zhou Y.H., Takekoshi M., Maeda F., Ihara S., Esumi M.: Recombinant antibody Fab against the hypervariable region 1 of hepatitis C virus blocks the virus adsorption to susceptible cellsin vitro.Antiviral Res. 56, 51–59 (2002).

    Article  PubMed  CAS  Google Scholar 

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  1. Institute of Microbiology and Immunology, Faculty of Veterinary Medicine, Veterinary and Pharmaceutical University, 612 42, Brno, Czechia

    D. Blažek & V. Celer

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  1. D. Blažek
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This work was supported by theGrant Agency of the Czech Republic (grant no. 524/01/0819).

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Blažek, D., Celer, V. The production and application of single-chain antibody fragments. Folia Microbiol 48, 687–698 (2003). https://doi.org/10.1007/BF02993480

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