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Human Antibody Fragments Specific for Human Blood Group Antigens from a Phage Display Library

Abstract

We have isolated human antibody fragments with binding specificities against the blood group antigens of the ABO and I blood group systems (B and HI), of the Rh system (D and E) and of the Kell system (Kpb), by selecting a phage-antibody library on human red cells. The fragments, expressed in bacteria, were antigen-specific and effective hi assays including agglutination and immunohistochemistry. Selection of phage-antibody libraries using intact cells seems to offer a promising alternative to hybridoma technology for the production of antibodies against cell surface molecules.

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References

  1. 1

    Winter, G. and Milstein, C. 1991. Man-made antibodies. Nature 349: 293–299.

    CAS  Article  Google Scholar 

  2. 2

    Marks, J.D., Hoogenboom, H.R., Bonnert, T.P., McCafferty, J., Griffiths, A.D. and Winter, G. 1991. By-passing immunization: Human antibodies from V-gene libraries displayed on phage. J. Mol. Biol. 222: 581–597.

    CAS  Article  Google Scholar 

  3. 3

    Griffiths, A.D., Malmqvist, M., Marks, J.D., Bye, J.M., Embleton, M.J., McCafferty, J., Baier, M., Holliger, K.P., Gorick, B.D., Hughes-Jones, N.C., Hoogenboom, H.R. and Winter, G. 1993. Human anti-self antibodies with high specificity from phage display libraries. EMBO J. 12: 725–734.

    CAS  Article  Google Scholar 

  4. 4

    Marks, J.D., Hoogenboom, H.R., Griffiths, A.D. and Winter, G. 1992. Molecular evolution of proteins on filamentous phage: mimicking the strategy of the immune system. J. Biol. Chem. 267: 16007–16010.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. 5

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

    CAS  Article  Google Scholar 

  6. 6

    Marks, J.D., Griffiths, A.D., Malmqvist, M., Clackson, T., Bye, J.M. and Winter, G. 1992. Bypassing immunization: high affinity human antibodies by chain shuffling. Bio/Technology 10: 779–783.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 7

    Hawkins, R., Russel, S.J. and Winter, G. 1992. Selection of phage antibodies by binding affinity: mimicking affinity maturation. J. Mol. Biol. 226: 889–896.

    CAS  Article  Google Scholar 

  8. 8

    Munro, S. and Pelham, H.R.B. 1986. An Hsp-like protein in the ER: Identity with the 78kd glucose regulated protein and immunoglobulin heavy chain binding protein. Cell 46: 291–300.

    CAS  Article  Google Scholar 

  9. 9

    Bye, J.M., Carter, C., Cui, Y., Gorick, B.D., Songsvilai, S., Winter, G., Hughes-Jones, N.C. and Marks, J.D. 1992. Germline variable region gene segment derivation of human monoclonal anti-Rh (D) antibodies. J. Clin. Invest. 90: 2481–2490.

    CAS  Article  Google Scholar 

  10. 10

    Parsons, S.F., Gardner, B. and Anstee, D.J. 1992. Monoclonal antibodies against Kelt glycoprotein: Serology, immunochemistry and quantification of antigen sites. Trans. Med. 3: 137–142.

    Article  Google Scholar 

  11. 11

    Cherif-Zahar, B., Bloy, C., Le Van Kim, C., Blanchard, D., Bailly, P., Hermand, P., Salmon, C., Cartron, J.-P. and Colin, I. 1990. Molecular cloning and protein structure of a human blood group Rh polypeptide. Proc. Natl. Acad. Sci. USA 87: 6243–6247.

    CAS  Article  Google Scholar 

  12. 12

    Mollison, P.L., Englefriet, C.P. and Contreras, M. 1993. Blood Transfusion in Clinical Medicine. Blackwell Scientific Publications, Oxford.

    Google Scholar 

  13. 13

    Voak, D., Lodge, T.W., Hopkins, J. and Bowley, C.C. 1968. A study of the antibodies of the HOI-B complex with special reference to their occurrence and notation. Vox Sang. 15: 353–366.

    CAS  PubMed  Google Scholar 

  14. 14

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

    CAS  Article  Google Scholar 

  15. 15

    Barbas, C.F., Collet, T.A., Amberg, W., Roben, P., Binley, J.M., Hoekstra, D., Cababa, D., Jones, T.M., Williamson, R.A., Pilkington, G.R., Haig-wood, N.L., Cabezas, E., Satterthwait, A.C., 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: 812–823.

    CAS  Article  Google Scholar 

  16. 16

    Miller, J.H. 1972. Experiments in Molecular Genetics. Cold Springs Harbor Lab Press. Cold Springs Harbor, New York.

    Google Scholar 

  17. 17

    Hoogenboom, H.R., Griffiths, A.D., Johnson, K.S., Chiswell, D.J., Hudson, P. and Winter, G. 1991. Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains. Nucl. Acids Res. 19: 4133–4137.

    CAS  Article  Google Scholar 

  18. 18

    De Bellis, D. and Schwartz, I. 1990. Regulated expression of foreign genes fused to lac: control by glucose levels in growth medium. Nucl. Acids Res. 18: 1311.

    CAS  Article  Google Scholar 

  19. 19

    Gussow, D. and Clackson, T. 1989. Direct clone characterization from plaques and colonies by the polymerase chain reaction. Nucl. Acids Res. 17: 4000.

    CAS  Article  Google Scholar 

  20. 20

    Sanger, F., Nicklen, F. and Coulson, A.R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463–5467.

    CAS  Article  Google Scholar 

  21. 21

    Tomlinson, I.M., Walter, G., Marks, J.D., Llewelyn, M.B. and Winter, G. 1992. The repertoire of human germline VH sequences reveals about fifty groups of VH segments with different hypervariable loops. J. Mol. Biol. 227: 776–798.

    CAS  Article  Google Scholar 

  22. 22

    Bentley, D.L. and Rabbits, T.H. 1980. Human immunoglobulin variable region genes—DNA sequences of two V kappa genes and a pseudogene. Nature 288: 730–733.

    CAS  Article  Google Scholar 

  23. 23

    Bentley, D.L. and Rabbitts, T.H. 1983. Evolution of immunoglobulin V genes: Evidence indicating that recently duplicated human V sequences have diverged by gene conversion. Cell 32: 181–189.

    CAS  Article  Google Scholar 

  24. 24

    Jaenichen, H.R., Pech, M., Lindenmaier, W., Wildgruber, N. and Zachau, H.G. 1984. Composite human VK genes and a model of their evolution. Nucl. Acids Res. 12: 5249–5263.

    CAS  Article  Google Scholar 

  25. 25

    Pech, M. and Zachau, H.G. 1984. Immunoglobulin genes of different subgroups are interdigitated within the VK locus. Nucl. Acids Res. 12: 9229–9236.

    CAS  Article  Google Scholar 

  26. 26

    Pech, M., Jaenichen, H.R., Pohlenz, H.D., Neumaier, P.S., Klobeck, H.G. and Zachau, H.G. 1984. Organization and evolution of a gene cluster for human immunoglobulin variable regions of the kappa type. J. Mol. Biol. 176: 189–204.

    CAS  Article  Google Scholar 

  27. 27

    Pech, M., Smola, H., Pohlenz, H.D., Straubinger, B., Gerl, R. and Zachau, H.G. 1985. A large section of the gene locus encoding human immunoglobulin variable regions of the kappa type is duplicated. J. Mol. Biol. 183: 291–299.

    CAS  Article  Google Scholar 

  28. 28

    Klobeck, H.G., Bornkamm, G., Combriato, W., Mocikat, R., Pohlenz, H.D. and Zachau, H.G. 1985. Subgroup IV of human immunoglobulin K light chains is encoded by a single gerrnline gene. Nucl. Acids Res. 13: 6515–6529.

    CAS  Article  Google Scholar 

  29. 29

    Klobeck, H.G., Meindl, A., Combriato, G., Solomon, A. and Zachau, H.G. 1985. Human immunoglobulin kappa light chain genes of subgroups II and III. Nucl. Acids Res. 13: 6499–6513.

    CAS  Article  Google Scholar 

  30. 30

    Stavnezer, J., Kekish, O., Batter, D., Grenier, J., Balazs, I., Henderson, E. and Zegers, B.J. 1985. Aberrant recombination events in B cell lines derived from a kappa-deficient human. Nucl. Acids Res. 13: 3495–3514.

    CAS  Article  Google Scholar 

  31. 31

    Chen, P.P., Albrandt, K., Orida, N.K., Radoux, V., Chen, E.Y., Schrantz, R., Liu, F.T. and Carson, D.A. 1986. Genetic basis for the cross-reactive idiotypes on the light chains of human IgM anti-IgG autoantibodies. Proc. Natl. Acad Sci. USA 83: 8318–8322.

    CAS  Article  Google Scholar 

  32. 32

    Chen, P.P., Robbins, D.L., Jirik, F.R., Kipps, T.J. and Carson, D.A. 1987. Isolation and characterization of a light chain variable region gene for human rheumatoid factors. J. Exp. Med. 166: 1900–1905.

    CAS  Article  Google Scholar 

  33. 33

    Chen, P.P., Albrandt, K., Kipps, T.J., Radoux, V., Liu, F.-T. and Carson, D.A. 1987. Isolation and characterization of human VkIII germ-line genes. Implications for the molecular basis of human VkIII light chain diversity. J. Immunol. 139: 1727–1733.

    CAS  PubMed  Google Scholar 

  34. 34

    Lorenz, W., Schable, K.F., Thiebe, R., Stavnezer, J. and Zachau, H.G. 1988. The J kappa proximal region of the human K locus contains three uncommon V Kappa genes which are arranged in opposite transcriptional polarities. Mol. Immunol. 25: 479–484.

    CAS  Article  Google Scholar 

  35. 35

    Straubinger, B., Huber, E., Osterholzer, E., Pargent, W., Pech, M., Pohlenz, H.D., Zimmer, F.J. and Zachau, H.G. 1988. The human VK locus. Characterization of a duplicated region encoding 28 different immunoglobulin genes. J. Mol. Biol. 199: 23–34.

    CAS  Article  Google Scholar 

  36. 36

    Straubinger, B., Thiebe, R., Huber, C., Osterholzer, E. and Zachau, H.G. G.1988. Two unusual human immunoglobulin V kappa genes. Biol. Chem. Hoppe Seyler 369: 601–607.

    CAS  Article  Google Scholar 

  37. 37

    Scott, M.G., Crimmins, D.L., McCourt, D.W., Zocher, I., Thiebe, R., Zachau, H.G. and Nahm, M.H. 1989. Clonal characterization of the human IgG antibody repertoire to Haemophilus influenzae type b polysaccharide. III. A single VKII gene and one of several JK genes are joined by an invariant arginine to form the most common L chain V region. J. Immunol. 143: 4110–4116.

    CAS  PubMed  Google Scholar 

  38. 38

    Scott, J.K. and Smith, G.P. 1990. Searching for peptide ligands with an epitope library. Science 249: 386–390.

    CAS  Article  Google Scholar 

  39. 39

    Pargent, W., Meindl, A., Thiebe, R., Mitzel, S. and Zachau, H.G. 1991. The human immunoglobulin kappa locus: characterization of the duplicated O regions. Eur. J. Immunol. 21: 1821–1827.

    CAS  Article  Google Scholar 

  40. 40

    Lautner-Rieske, A., Huber, C., Meindle, A., Pargent, W., Schable, K.F., Thiebe, R., Zocher, I. and Zachau, H.G. 1991. The human immunoglobulin kappa locus: characterization of the dublicated A regions. Eur. J. Immunol. 22: 1023–1029.

    Article  Google Scholar 

  41. 41

    Williams, S.C. and Winter, G. 1993. Cloning and sequencing of human immunoglobulin variable lambda gene segments. Eur. J. Immunol. 23: 1456–1461.

    CAS  Article  Google Scholar 

  42. 42

    Ward, E.S., Gussow, D., Griffiths, A.D., Jones, P.T. and Winter, G. . 1989. Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature 341: 544–546.

    CAS  Article  Google Scholar 

  43. 43

    Thorpe, S.J. 1989. Detection of Rh D-associated epitopes in human and animal tissues using human\monoclonal anti-D antibodies. Brit. J. Haemal 73: 527–536.

    CAS  Article  Google Scholar 

  44. 44

    Frippiat, J.-P., Chuchuna, P., Bernard, F., Bulewala, L., Lefranc, G. and Lefranc, J.-P. 1990. First genomic sequence of a human Ig variable lambda gene belonging to subgroup III. Nucl. Acids Res. 18: 7134.

    CAS  Article  Google Scholar 

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Marks, J., Ouwehand, W., Bye, J. et al. Human Antibody Fragments Specific for Human Blood Group Antigens from a Phage Display Library. Nat Biotechnol 11, 1145–1149 (1993). https://doi.org/10.1038/nbt1093-1145

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