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Synthetic Customized scFv Libraries

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Antibody Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 907))

Abstract

Libraries of antibody fragments displayed on filamentous phages have proved their value to generate human antibodies against virtually any target. We describe here a simple protocol to make large and diverse libraries based on a single or few frameworks. Diversity is introduced in the third hypervariable loops using degenerate synthetic oligonucleotides and PCR assembly. Because all the antibody fragments isolated from the library will share their framework sequence, their stability and physical properties will be more consistent and customizable than when antibody fragments are isolated from a library prepared from human donors.

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References

  1. Scolnik PA (2009) mAbs: a business perspective. MAbs 1:179–184

    Article  PubMed Central  PubMed  Google Scholar 

  2. Schofield DJ, Pope AR, Clementel V, Buckell J, Chapple SD, Clarke KF, Conquer JS, Crofts AM, Crowther SRE, Dyson MR, Flack G, Griffin GJ, Hooks Y, Howat WJ, Kolb-Kokocinski A, Kunze S, Martin CD, Maslen GL, Mitchell JN, O’Sullivan M, Perera RL, Roake W, Shadbolt SP, Vincent KJ, Warford A, Wilson WE, Xie J, Young JL, McCafferty J (2007) Application of phage display to high throughput antibody generation and characterization. Genome Biol 8:R254

    Article  PubMed Central  PubMed  Google Scholar 

  3. Demarest SJ, Glaser SM (2008) Antibody therapeutics, antibody engineering, and the merits of protein stability. Curr Opin Drug Discov Devel 11:675–687

    CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  5. Philibert P, Stoessel A, Wang W, Sibler A, Bec N, Larroque C, Saven JG, Courtete J, Weiss E, Martineau P (2007) A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm. BMC Biotechnol 7:81

    Article  PubMed Central  PubMed  Google Scholar 

  6. Silacci M, Brack S, Schirru G, Marlind J, Ettorre A, Merlo A, Viti F, Neri D (2005) Design, construction, and characterization of a large synthetic human antibody phage display library. Proteomics 5:2340–2350

    Article  CAS  PubMed  Google Scholar 

  7. Martineau P, Jones P, Winter G (1998) Expression of an antibody fragment at high levels in the bacterial cytoplasm. J Mol Biol 280:117–127

    Article  CAS  PubMed  Google Scholar 

  8. Sidhu SS, Lowman HB, Cunningham BC, Wells JA (2000) Phage display for selection of novel binding peptides. Methods Enzymol 328:333–363

    CAS  PubMed  Google Scholar 

  9. Ravn U, Gueneau F, Baerlocher L, Osteras M, Desmurs M, Malinge P, Magistrelli G, Farinelli L, Kosco-Vilbois M, Fischer N (2010) By-passing in vitro screening—next generation sequencing technologies applied to antibody display and in silico candidate selection. Nucleic Acids Res 38:e193–e193

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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Correspondence to Pierre Martineau .

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© 2012 Springer Science+Business Media, LLC

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Robin, G., Martineau, P. (2012). Synthetic Customized scFv Libraries. In: Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, vol 907. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-974-7_6

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  • DOI: https://doi.org/10.1007/978-1-61779-974-7_6

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-973-0

  • Online ISBN: 978-1-61779-974-7

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