A major ‘post-genome’ objective is to fully characterise the human proteome using a comprehensive collection of specific antibodies directed against all human proteins and their variants. To achieve this, substantial reductions in cost and effort per antibody have to be achieved. A conseqeunt use of microtitre plates amneable to automatisation is a prerequisite to produce antibodies in "proteome scale" numbers. The method decribed in this chapter allows the generation of antibodies by phage display with minimized effort and cost.
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References
Breitling F, Dübel S, Seehaus T, Klewinghaus I, Little M (1991) A surface expression vector for antibody screening. Gene 104:147–153
Finnern R, Pedrollo E, Fisch I, Wieslander J, Marks JD, Lockwood CM, Ouwehand WH (1997) Human autoimmune anti-proteinase 3 scFv from a phage display library. Clin Exp Immunol 107:269–281
Goletz S, Christensen PA, Kristensen P, Blohm D, Tomlinson I, Winter G, Karsten U (2002) Selection of large diversities of antiidiotypic antibody fragments by phage display. J Mol Biol 315:1087–1097
Hawlisch H, Müller M, Frank R, Bautsch W, Klos A, Köhl J (2001) Site-specific anti-C3a receptor single-chain antibodies selected by differential panning on cellulose sheets. Anal Biochem 293:142–145
Hust M, Maiss E, Jacobsen H, Reinard T (2002) The production of a genus-specific recombinant antibody (scFv) using a recombinant potyvirus protease. J Virol Methods 106:225–233
Hust M, Meysing M, Schirrmann T, Selke M, Meens J, Gerlach G, Dübel S (2006) Enrichment of open reading frames presented on bacteriophage M13 using hyperphage. Biotechniques 41:335–342
Hust M, Jostock T, Menzel C, Voedisch B, Mohr A, Brenneis M, Kirsch MI, Meier D, Dübel S (2007a) Single chain Fab (scFab) fragment. BMC Biotechnol 7:14
Hust M, Toleikis L, Dübel S (2007b) Antibody phage display. In Handbook of therapeutic antibodies. Wiley-VCH, Weinheim
Kirsch M, Hülseweh B, Nacke C, Rülker T, Schirrmann T, Marschall H, Hust M, Dübel S (2008) Development of human antibody fragments using antibody phage display for the detection and diagnosis of Venezuelan equine encephalitis virus (VEEV). BMC Biotechnol 8:66
Mersmann M et al (1998) Monitoring of scFv selected by phage display using detection of scFv-pIII fusion proteins in a microtiter scale assay. J Immunol Methods 220:51–58
Moghaddam A, Borgen T, Stacy J, Kausmally L, Simonsen B, Marvik OJ, Brekke OH, Braunagel M (2003) Identification of scFv antibody fragments that specifically recognise the heroin metabolite 6-monoacetylmorphine but not morphine. J Immunol Methods 280:139–155
Parmley SF, Smith GP (1988) Antibody-selectable filamentous fd phage vectors: affinity purification of target genes. Gene 73:305–318
Pelat T, Hust M, Laffly E, Condemine F, Bottex C, Vidal D, Lefranc M, Dübel S, Thullier P (2007) High-affinity, human antibody-like antibody fragment (single-chain variable fragment) neutralizing the lethal factor (LF) of Bacillus anthracis by inhibiting protective antigen-LF complex formation. Antimicrob Agents Chemother 51:2758–2764
Retter I, Althaus HH, Münch R, Müller W (2005) VBASE2, an integrative V gene database. Nucleic Acids Res 33:D671–D674
Rondot S, Koch J, Breitling F, Dübel S (2001) A helper phage to improve single-chain antibody presentation in phage display. Nat Biotechnol 19:75–78
Soltes G, Hust M, Ng KKY, Bansal A, Field J, Stewart DIH, Dübel S, Cha S, Wiersma EJ (2007) On the influence of vector design on antibody phage display. J Biotechnol 127:626–637
Welschof M, Terness P, Kipriyanov SM, Stanescu D, Breitling F, Dörsam H, Dübel S, Little M, Opelz G (1997) The antigen-binding domain of a human IgG-anti-F(ab’)2 autoantibody. Proc Natl Acad Sci USA 94:1902–1907
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Hust, M., Mersmann, M. (2010). Phage Display and Selection in Microtitre Plates. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-01144-3_10
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