Advertisement

Generation of Naive Human Antibody Libraries

Part of the Springer Lab Manuals book series (SLM)

Abstract

The phage display of antibody fragments as Fabs or scFvs (McCafferty et al., 1990), has its origins in experiments demonstrating that both small peptides and folded proteins could be displayed on the surface of filamentous bacteriophage (Smith, 1985; Bass et al., 1990). Since the generation of the first human antibodies by phage display (Winter et al., 1994), the technology has developed to the point where large scFv repertoires have been created that yield antibodies with sub-nanomolar affinities (Vaughan et al., 1996; Xie et al., 1997). This is comparable with the best antibodies obtained using hybridoma technology. Phage display repertoires can also be used to isolate antibodies not easily obtained by hybridoma technology, such as those specific for toxic proteins and human anti-self antibodies (Griffiths et al., 1993; Vaughan et al., 1998). In addition, using a variety of selection and screening strategies, the same non-immunised “single pot“ library can be used to simultaneously derive many high affinity antibodies with different specificities in only a matter of weeks. Furthermore, selection techniques have evolved to the point where strategies now exist that facilitate selections on complex antigens expressed on the cell surface (Chapter 12).

Keywords

Phage Display Phage Stock scFv Library Phagemid Particle Clone scFv 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bass S, Greene R and Wells JA (1990) Hormone phage: an enrichment method for variant proteins with altered binding properties. Proteins 8:309–314PubMedCrossRefGoogle Scholar
  2. Glover DR, (1999) Fully human antibodies come to fruition. SCRIP Magazine (May): 16–19Google Scholar
  3. Griffiths AD, Malmqvist M, Marks JD, Bye JM, Embleton MJ, McCafferty J et al (1993) Human anti-self antibodies with high specificity from phage display libraries. EMBO J. 12:725–734PubMedGoogle Scholar
  4. Jespers LS, Roberts A, Mahler SM, Winter G and Hoogenboom HR (1994) Guiding the selection of human antibodies from phage display repertoires to a single epitope of an antigen. BioTechnology 12: 899–903PubMedCrossRefGoogle Scholar
  5. Johnson KS and Glover DR (1998) Antibodies come in from the cold. Innovations in Pharmaceutical Technology p82–85Google Scholar
  6. Marks JD, Hoogenboom HR, Bonnert TP, McCafferty J, Griffiths AD and Winter G (1991) By-passing immunization: Human antibodies from V-gene libraries displayed on phage. J Mol Biol 222:581–597PubMedCrossRefGoogle Scholar
  7. McCafferty J, Griffiths AD, Winter G and Chiswell D (1990) Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348:552–554PubMedCrossRefGoogle Scholar
  8. Sambrook J, Fritsch EF and Maniatis T (1990) Molecular cloning — a laboratory manual. Cold Spring Harbor Laboratory. New York.Google Scholar
  9. Smith GP (1985) Filamentous phage: novel expression vectors that display cloned antigens on the virion surface. Science 228(4705): 1315–1317PubMedCrossRefGoogle Scholar
  10. Vaughan TJ, Williams AJ, Pritchard K, Osbourn JK, Pope AR, Earnshaw JC, McCafferty J, Hodits RA, Wilton J and Johnson KS (1996) Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library. Nature Biotechnology 14: 309–314PubMedCrossRefGoogle Scholar
  11. Vaughan TJ, Osbourn JK and Tempest PR (1998) Human antibodies by design. Nature Biotechnology 16: 535–539PubMedCrossRefGoogle Scholar
  12. Xie M-H, Yuan J, Adams C and Gurney A (1997) Direct demonstration of MuSK involvement in acetylcholine receptor clustering through identification of agonist scFv. Nature Biotechnology 15:768–771PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2001

Authors and Affiliations

  • Catherine Hutchings
    • 1
  • Sara Carmen
    • 1
  • Simon Lennard
    • 1
  1. 1.Cambridge Antibody TechnologyMelbourn, CambridgeshireUK

Personalised recommendations