Phage-Display Libraries

  • Julia E. Thompson
  • Andrew J. Williams
Part of the Springer Protocols Handbooks book series (SPH)


Foreign proteins can be displayed on the surface of filamentous bacteriophage by fusing them to the phage minor coat protein (Fig. 1) Many proteins have been expressed in this way, including human growth factor (1), alkaline phosphatase (2), ricin (3), pancreatic trypsin inhibitor (4), and a DNA-binding protein (5) Most importantly, the displayed proteins retain their biological activity


Phage Display Antibody Fragment Phage Particle Human Neutrophil Elastase Filamentous Bacteriophage 
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  1. 1.
    Bass, S, Greene, R, and Wells, J A. (1990) Hormone phage an enrichment method for variant proteins with altered binding properties Proteins 8, 309–314PubMedCrossRefGoogle Scholar
  2. 2.
    McCafferty, J, Jackson, R H, and Chiswell, D J (1991) Phage-enzymes expression and affinity chromatography of functional alkaline phosphatase on the surface of bacte-nophage Protein Eng 4, 955–961PubMedCrossRefGoogle Scholar
  3. 3.
    Swimmer, C, Lehar, S M, McCafferty, J, Chiswell, D J, Blattler, W A, and Guild, B (1992) Phage display of ricin chain and its single binding domains system for screening galactose-binding mutants Proc Natl Acad Sa USA 89, 3756–3760CrossRefGoogle Scholar
  4. 4.
    Roberts, L, Markland, W, Ley,, Kent, R, White, D W, Guterman, S, and Ladner, R (1992) Directed evolution of a protein selection of potent neutrophil elastase inhibitors displayed on Ml3 fusion phage Proc Natl Acad Sei USA 89, 2429–2433CrossRefGoogle Scholar
  5. 5.
    Rebar, E J and Pabo, C O (1994) Zmc finger phage affinity selection of fingers with new DNA-binding specificities Science 263, 671–673PubMedCrossRefGoogle Scholar
  6. 6.
    Winter, G, Griffiths, A D, Hawkins, R E, and Hoogenboom, H R (1994) Making antibodies by phage display technology Annu Rev Immunol 12, 433–455Google Scholar
  7. 7.
    Clackson, T and Wells, J A (1994) In vitro selection from protein and peptide libraries Trends Bwtechnol 12, 173–184CrossRefGoogle Scholar
  8. 8.
    Kohler, G and Mustern, C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity Nature (Lond) 256, 495–497CrossRefGoogle Scholar
  9. 9.
    Winter, G and Milstein, C (1991) Man-made antibodies Nature (Lond) 349, 293–299CrossRefGoogle Scholar
  10. 10.
    LoBugho, A F, Wheeler, R H, Trang, J, Haynes, A, Rogers, K, Harvey, E, Sun, L, Ghrayeb, J, and Khazeli, M (1989) Mouse/human chimaeric monoclonal antibody in man kinetics and immune responses Proc Natl Acad Sei USA 86, 4220–4224CrossRefGoogle Scholar
  11. 11.
    James, K and Bell, G T (1987) Human monoclonal antibody production current status and future prospects J Immunol Meth 100, 5–40CrossRefGoogle Scholar
  12. 12.
    Nossal, G. J (1989) Immunologie tolerance collaboration between antigen and lymphok-mes Science 245, 147–153PubMedCrossRefGoogle Scholar
  13. 13.
    Larrick, J W, Danielsson, L, Brenner, A, Abrahamson, M, Fry, K E, and Borrebaeck, C A (1989) Rapid cloning of rearranged immunoglobulin genes from human hybridoma cells using mixed primers and the polymerase chain reaction Biochem Biophys Res Commun 160, 1250–1256PubMedCrossRefGoogle Scholar
  14. 14.
    Mulhnax, R L., Gross, E A, Amberg, J R, Hay, N, Hogrefe, H H, Kubitz, M M, Greener, A, Alting, M M, Ardourel, D, Short, J M, Sorge, J A, and Shopes, B (1990) Identification of a human antibody fragment clones specific for tetanus toxoid in a bacte-riophage lambda immunoexpression library Proc Natl Acad Sei USA 87, 8095–8099CrossRefGoogle Scholar
  15. 15.
    Persson, M A A, Caothien, R H, and Burton, D R (1991) Generation of diverse high-affinity human monoclonal antibodies by repertoire cloning Proc Natl Acad Sei USA 88, 2432–2436CrossRefGoogle Scholar
  16. 16.
    Marks, J D, Tnstrem, M, Karpas, A, and Winter, G (1991) Oligonucleotide primers for polymerase chain reaction amplification of human immunoglobulin variable genes and design of family-specific oligonucleotide probes Eur J Immunol 21, 985–991PubMedCrossRefGoogle Scholar
  17. 17.
    McCafferty, J, Griffiths, A D, Winter, G, and Chiswell, D J (1990) Phage antibodies filamentous phage displaying antibody variable domains Nature (Lond) 348, 552–554CrossRefGoogle Scholar
  18. 18.
    Hoogenboom, H R, Griffiths, A D, Johnson, K S, Chiswell, D J, Hudson, P, and Winter, G (1991) Multisubunit proteins on the surface of filamentous phage methodologies for displaying antibody (Fab) heavy and light chains Nucleic Acids Res 19, 4133–4137PubMedCrossRefGoogle Scholar
  19. 19.
    Barbas, F, Kang, A S, Lerner, R A, and Benkovic, S J (1991) Assembly of combinatorial antibody libraries on phage surfaces the gene III site Proc Natl Acad Sei USA 88, 7978–7982CrossRefGoogle Scholar
  20. 20.
    Better, M, Chang, P, Robinson, R R, and Horwitz, A H (1988) Escherichia coli secretion of an active chimenc antibody fragment Science 240, 1041–1043PubMedCrossRefGoogle Scholar
  21. 21.
    Skera, A and Pluckthun, A. (1988) Assembly of a functional immunoglobulin Fv fragment in Escherichia coli Science 240, 1038–1041Google Scholar
  22. 22.
    Marks, J D, Hoogenboom, H R, Bonnert, T P, McCafferty, J, Griffiths, A D, and Winter, G (1991) By-passing immunisation human antibodies from V-gene libraries displayed on phage J Mol Biol 222, 581–597PubMedCrossRefGoogle Scholar
  23. 23.
    Griffiths, A D, Malmqvist, M, Marks, J D, Bye, J M, Embleton, M J, McCafferty, J, Baier, M, Holliger, K P, Gonck, D, Hughes-Jones, N, Hoogenboom, H R, and Winter, G (1993) Human anti-self antibodies with high specificity from phage display libraries EMBO J 12, 725–734PubMedGoogle Scholar
  24. 24.
    de Kruif, J, Boel, E, and Logtenberg, T (1995) Selection and application of human single chain Fv antibody fragments from a semi-synthetic phage antibody display library with designed CDR3 regions J Mol Biol 248, 97–105PubMedCrossRefGoogle Scholar
  25. 25.
    Caton, A J and Koprowski, H (1990) Influenza virus hemagglutinin-specific antibodies isolated from a combinatorial expression library are closely related to the immune response of the donor Proc Natl Acad Sa USA 87, 6450–6454CrossRefGoogle Scholar
  26. 26.
    Lerner, R A, Barbas, R, Kang, A S, and Burton, D R (1991) On the use of combinatorial antibody libraries to clone the “fossil record” of an individual’s immune response Proc Natl Acad Sa USA 88, 9705,9706CrossRefGoogle Scholar
  27. 27.
    Barbas, F, Crowe, J E, Cababa, D, Jones, T M, Zebedee, S L, Murphy, R, Chanock, R M, and Burton, D R (1992) Human monoclonal Fab fragments derived from a combinatorial library bind to respiratory syncytial virus F glycoprotein and neutralise infectivity Proc Natl Acad Sei USA 89, 10,164–10,168CrossRefGoogle Scholar
  28. 28.
    Williamson, R A, Bunoni, R, Sanna, P P, and Partridge, L J (1993) Human monoclonal antibodies against a plethora of viral pathogens from single combinatorial libraries Proc Natl Acad Sei USA 90, 4141–4145CrossRefGoogle Scholar
  29. 29.
    Zebedee, S L Barbas, F, Horn, Y-L, Caothien, R H, Graff, R, DeGraw, J, Pyati, J, LaPolla, R, Burton, D R, Lerner, R A, and Thornton, G (1992) Human combinatorial antibody libraries to hepatitis surface antigen Proc Natl Acad Sei USA 89, 3175–3179CrossRefGoogle Scholar
  30. 30.
    Rapoport, B, Portolano, S, and McLachlan, S M (1995) Combinatorial libraries new insights into human organ-specific autoantibodies Immunol Today 16, 43–49PubMedCrossRefGoogle Scholar
  31. 31.
    Barbas, F., Collet, T A, Amberg, W, Roben, P, Binley, J M, Hoekstra, D, Cababa, D, Jones, T M, Williamson, R A, Pilkington, G R, Haingwood, N L, Cabezas, E, Satterthwait, A, Sanz, I, and Burton, D R (1993) Molecular profile of an antibody response to HIV-1 as probed by combinatorial libraries J Mol Biol 230, 812–823PubMedCrossRefGoogle Scholar
  32. 32.
    Vaughan, T J, Williams, A J, Pritchard, K, Osbourn, J K., Pope, A R, Earashaw, J, McCafferty, J, and Johnson, K S (1995) Isolation of human antibodies with sub-nanomolar affinities directly from a large non-immunized phage display library Biotechnology, submittedGoogle Scholar
  33. 33.
    Griffiths, A D, Williams, S, Hartley,, Tomlinson, I M, Waterhouse, P, Crosby, W L, Kontermann, R E, Jones, P T, Low, N M, Allison, T J, Prospera, T D, Hoogenboom, H R, Nissim, A, Cox, J P L, Harrison, J L, Zaccolo, M, Gherardi, E, and Winter, G (1994) Isolation of high affinity human antibodies directly from large synthetic repertoires EMBO J 13, 3245–3260PubMedGoogle Scholar
  34. 34.
    Vieira, J and Messing, J (1987) Production of single-stranded plasmid DNA Meth Enzymol 153, 3–11PubMedCrossRefGoogle Scholar
  35. 35.
    Garrard, L J, Yang, M, O’Connell, M P, Kelley, R F, and Henner, D J (1991) Fab assembly and enrichment in a monovalent phage display system Biotechnology 9, 1373–1377PubMedCrossRefGoogle Scholar
  36. 36.
    Lowman, H, Bass, S H, Simpson, N, and Wells, J A (1991) Selecting high-affinity binding proteins by monovalent phage display Biochemistry 30, 10,832–10,838PubMedCrossRefGoogle Scholar
  37. 37.
    Clackson, T, Hoogenboom, H R, Griffiths, A D, and Winter, G (1991) Making antibody fragments using phage display libraries Nature (Lond) 352, 624–628CrossRefGoogle Scholar
  38. 38.
    Hawkins, R E, Russell, S J, and Winter, G (1992) Selection of phage antibodies by binding affinity. Mimicking affinity maturation J Mol Biol 226, 889–896PubMedCrossRefGoogle Scholar
  39. 39.
    Thompson, J E, Pope, A R, Tung, J-S, Chan, C, Hollis, G, Mark, G, and Johnson, K S (1995) Affinity maturation of a high affinity human monoclonal antibody against the third hypervanabie loop of Human Immunodeficiency Virus use of phage display to improve affinity and broaden strain reactivity J Mol Biol, submittedGoogle Scholar
  40. 40.
    Barbas, F, Hu, D, Dunlop, N, Sawyer, L, Dababa, D, Hendry, R M, Nara, P L, and Burton, D R (1994) In vitro evolution of a neutralizing human antibody to human immunodeficiency virus type 1 to enhance affinity and broaden strain cross-reactivity Proc Natl Acad Sei USA 91, 3809–3813CrossRefGoogle Scholar
  41. 41.
    Kang, A S, Jones, T M, and Burton, D R (1991) Antibody redesign by chain shuffling from random combinatorial immunoglobulin libraries Proc Natl Acad Sci USA 88, 11,120–11,123PubMedCrossRefGoogle Scholar
  42. 42.
    Leung, D W, Chen, E, and Goeddel, D V (1989) A method for random mutagenesis of a defined DNA segment using a modified polymerase chain reaction Technmque 1, 11–15Google Scholar
  43. 43.
    Foote, J and Eisen, H N (1995) Kinetic and affinity limits on antibodies produced during immune responses Proc Natl Acad Sei USA 92, 1254–1256CrossRefGoogle Scholar
  44. 44.
    Huston, J S, McCartney, J, Tai, M-S, Mottola-Hartshorn,, Jin, D, Warren, F, Keck, P, and Oppermann, H (1993) Medical applications of single-chain antibodies Intern Rev Immunol 10, 195–217CrossRefGoogle Scholar
  45. 45.
    Lmsley, P S, Ledbetter, J A, Kinney-Thomas, E, and Hu, S L (1988) Effects of anti-gpl20 monoclonal antibodies on CD4 receptor binding by the env protein of human immunodeficiency virus type I J Virol 62, 3695–3702Google Scholar
  46. 46.
    Skinner, M A, Langlois, A J, McDanal, C B, McDougal, J S, Bolognesi, D P, and Matthews, T J (1988) Neutralising antibodies to an immunodominant envelope sequence do not prevent gpl20 binding to CD4 J Virol 62, 4195–4200PubMedGoogle Scholar
  47. 47.
    Emim, E A, Schleif, W A, Nunberg, J H., Conley, A J, Eda, Y, Tokiyoshi, S, Putney, S D, Matsushita, S, Cobb, E, Jett, M, Eichberg, J W, and Murthy, K K (1992) Prevention of HIV-1 infection in chimpanzees by gpl20 V3 domain-specific monoclonal antibody Nature (Lond) 355, 728–730CrossRefGoogle Scholar
  48. 48.
    Marks, J D, Griffiths, A D, Malmqvist, M, Clackson, T, Bye, J M, and Winter, G (1992) By-passing immunization building high affinity human antibodies by chain shuffling Bio/Technology 10, 779–783PubMedCrossRefGoogle Scholar
  49. 49.
    Ditzel, H J, Binley, J M, Moore, J P, Sodroski, J, Sullivan, N, Sawyer, L S W, Hendry, R M, Yang, W-P, Barbas, F, and Burton, D R. (1995) Neutralizing recombinant human antibodies to a conformational V-2 and CD4-binding site-sensitive epitope of HIV-1 gp 120 isolated by using an epitope masking procedure J Immunol 154, 893–906PubMedGoogle Scholar
  50. 50.
    Cook, B. and Beastall, G (1989) Steroid hormones a practical approach In Practical Approach Series (Rickwood, D and Hames, B. D, eds), Oxford University Press, Oxford, UK, pp 1–65Google Scholar
  51. 51.
    Pope, A R, Pntchard, K, Williams, A J, Hackett, J R, Roberts, A, Mandecki, W, and Johnson, K S (1995) In vitro selection of a high affinity antibody to oestradiol using a phage display human antibody library J Mol Biol, submittedGoogle Scholar
  52. 52.
    Figini, M, Marks, J D, Winter, G, and Griffiths, A D (1994) In vitro assembly of repertoires of antibody chains by renaturation on the surface of phage J Mol Biol 239, 68–78PubMedCrossRefGoogle Scholar
  53. 53.
    Jespers, L S, Roberts, A, Mahler, S M, Winter, G, and Hoogenboom, H R (1994) Guiding the selection of human antibodies from phage display repertoires to a single epitope of an antigen Bio/Technology 12, 899–903PubMedCrossRefGoogle Scholar
  54. 54.
    Barbas, F (1993) Recent advances in phage display Curr Opin Biotechnol 4, 526–530PubMedCrossRefGoogle Scholar
  55. 55.
    Lowman, H and Wells, J A (1993) Affinity maturation of human growth hormone by monovalent phage display J Mol Biol 234, 564–578PubMedCrossRefGoogle Scholar
  56. 56.
    Smith, G P (1985) Filamentous fusion phage novel expression vectors that display cloned antigens on the surface of the vinon Science 228, 1315–1317PubMedCrossRefGoogle Scholar
  57. 57.
    Scott, J and Smith, G P (1990) Searching for peptide hgands with an epitope library Science 249, 386–390PubMedCrossRefGoogle Scholar
  58. 58.
    Devlin, J J, Panganiban, L, and Devlin, P E (1990) Random peptide libraries, a source of specific protein binding molecules Science 249, 386–390CrossRefGoogle Scholar
  59. 59.
    Lane, D P and Stephen, W (1993) Epitope mapping usmg bactenophage peptide libraries Curr Opin Immunol 5, 268–271PubMedCrossRefGoogle Scholar
  60. 60.
    Matthews, D J and Wells, J A (1993) Substrate phage, selection of protease substrates by monovalent phage display Science 260, 1113–1117PubMedCrossRefGoogle Scholar
  61. 61.
    Scott, J K, Loganathan, D, Easley, R, Gong, X, and Goldstein, I J (1992) A family of concanavahn A-bindmg peptides from a hexapeptide epitope library Proc Natl Acad Sei USA 89, 5398–5402CrossRefGoogle Scholar
  62. 62.
    Oldenburg, K R, Loganathan, D, Goldstein, I J, Schultz, P G, and Gallop, M A (1992) Peptide hgands for a sugar-binding protein isolated from a random peptide library Proc Natl Acad Sa USA 89, 5393–5397CrossRefGoogle Scholar
  63. 63.
    O’Neil, K T, Hoess, R. H, Jackson, S A, Ramachandran, N S, Mousa, S A, and DeGrado, W F (1992) Identification of novel peptide antagonists for GPIIb/IIa from a conformationally constrained phage peptide library Proteins 14, 509–515PubMedCrossRefGoogle Scholar
  64. 64.
    Barbas, F, Bain, J D, Hoekstra, D M, and Lerner, R A (1992) Semisynthetic combinatorial antibody libraries a chemical solution to the diversity problem Proc Natl Acad Sa USA 89, 4457–4461CrossRefGoogle Scholar
  65. 65.
    Hoogenboom, H R and Winter, G (1992) By-passing immunization human antibodies from synthetic repertoires of germlme VH gene segments rearranged in vitro J Mol Biol 227, 381–388PubMedCrossRefGoogle Scholar
  66. 66.
    Barbas, F, Bjorhng, E, Chioki, F, Dunlop, N., Cabab, D, Jones, T M, Zebedee, S L, Persson, M A A, Nara, P L, Norrby, E, and Burton, D R (1992) Recombmant human Fab fragments neutralize human type 1 immunodeficiency virus in vitro Proc Natl Acad Sei USA 89, 9339–9343CrossRefGoogle Scholar
  67. 67.
    Nissim, A, Hoogenboom, H R, Tomhnson, I M, Flynn, G, Midgley, C, Lane, D, and Winter, G (1994) Antibody fragments from a’ single pot’ phage display library as immu-nochemical reagents EMBO J 13, 692–698PubMedGoogle Scholar

Copyright information

© Humana Press Inc , Totowa, NJ. 1998

Authors and Affiliations

  • Julia E. Thompson
    • 1
  • Andrew J. Williams
    • 1
  1. 1.Cambridge Antibody Technology, IncUK

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