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6 - Generation and Screening of the Synthetic Human Combinatorial Antibody Library HuCAL GOLD

from PART II - GENERATION AND SCREENING OF ANTIBODY LIBRARIES

Published online by Cambridge University Press:  15 December 2009

By
Ingo M. Klagge
Edited by
Melvyn Little
Melvyn Little
Affiliation:
Affimed Therapeutics AG
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Summary

Nowadays, monoclonal antibodies are the fastest growing class of biopharmaceuticals. By the end of 2007, the U.S. Food and Drug Administration (FDA) had approved 21 therapeutic antibodies. Since 1975, with the seminal work of Köhler and Milstein (Köhler & Milstein,1975) describing the use of hybridoma technology for monoclonal antibody generation, major advances in the field allowed for the development of antibody libraries using recombinant technologies (reviewed by Hoogenboom, 2005, and Sergeeva et al., 2006). Various display technologies and the integration of automated screening methods now enable researchers to quickly identify multiple target specific antibodies for later development as biopharmaceuticals.

This chapter will look at MorphoSys's latest fully human antibody library, the Human Combinatorial Antibody Library HuCAL GOLD based on phage display of Fab antibody fragments. Besides a comprehensive introduction of the design and generation of the library, the chapter will describe the HuCAL-specific CysDisplay technology, explore the use of MorphoSys's proprietary AgX technology, and give some examples on the use of HuCAL-based antibody optimization by using standard affinity maturation approaches or the recently developed RapMAT technology.

HuCAL CONCEPT

The HuCAL technology is a unique and innovative concept for the in vitro generation of highly diverse fully human antibodies. The structural basis for the HuCAL libraries is provided by seven heavy chain and seven light chain variable region genes (Knappik et al., 2000).

Type
Chapter
Information
Recombinant Antibodies for Immunotherapy , pp. 75 - 86
Publisher: Cambridge University Press
Print publication year: 2009

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References

Barbas, C.F., Hu, D., Dunlop, N., Sawyer, L., Cababa, D., Hendry, R.M., Nara., P.L., 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. Proceedings of the National Academy of Sciences USA, 91, 3809–18.CrossRefGoogle ScholarPubMed
Brocks, B., Kraft, S., Zahn, S., Noll, S., Pan, C., Schauer, M., Krebs, B. (2006). Generation and optimization of human antagonistic antibodies against TIMP-1 as potential therapeutic agents in fibrotic diseases. Human Antibodies, 15, 115–24.CrossRefGoogle ScholarPubMed
Ewert, S., Honegger, A., Plückthun, A. (2004). Stability improvement of antibodies for extracellular and intracellular applications: CDR grafting to stable frameworks and structure-based framework engineering. Methods, 34, 184–99.CrossRefGoogle ScholarPubMed
Frisch, C., Brocks, B., Ostendorp, R., Hoess, A., Rüden, T., Kretzschmar, T. (2003). From EST to ICH: human antibody pipeline for target research. Journal of Immunological Methods, 75, 203–12.CrossRefGoogle Scholar
Haenel, C., Satzger, M., Della Ducata, D., Ostendorp, R., Brocks, B. (2005). Characterization of high-affinity antibodies by electrochemiluminescence-based equilibrium titration. Analytical Biochemistry, 339, 182–4.CrossRefGoogle ScholarPubMed
Hillig, R.C., Urlinger, S., Fanghänel, J., Brocks, B., Haenel, C., Stark, Y., Sülzle, D., Svergun, D.I., Baesler, S., Malawski, G., Moosmayer, D., Menrad, A., Schirner, M., Licha, K. (2008). Fab MOR03268 triggers absorption shift of a diagnostic dye via packaging in a solvent-shielded Fab dimer interface. Journal of Molecular Biology, 377, 206–19.CrossRefGoogle Scholar
Hillig, R.C., Baesler, S., Urlinger, S., Stark, Y., Bauer, S., Badock, V., Huber, M., Bahr, I., Schirner, M., Licha, K. (2007). Crystallization and molecular-replacement solution of a diagnostic fluorescent dye in complex with a specific Fab fragment. Acta Crystallographica Section F, 63, 217–23.Google ScholarPubMed
Hoogenboom, H.R. (2005). Selecting and screening recombinant antibody libraries. Nature Biotechnology, 23, 1105–16.CrossRefGoogle ScholarPubMed
Jarutat, T., Nickels, C., Frisch, C., Stellmacher, F., Hofig, K.P., Knappik, A., Merz, H. (2007). Selection of vimentin-specific antibodies from the HuCAL phage display library by subtractive panning on formalin-fixed, paraffin-embedded tissue. Biological Chemistry, 388, 651–8.CrossRefGoogle ScholarPubMed
Knappik, A., Ge, L., Honegger, A., Pack, P., Fischer, M., Wellnhofer, G., Hoess, A., Wölle, J., Plückthun, A., Virnekäs, B. (2000). Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides. Journal of Molecular Biology, 296, 57–86.CrossRefGoogle ScholarPubMed
Köhler, G. and Milstein, C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, 256, 495–7.CrossRefGoogle ScholarPubMed
Krebs, B., Rauchenberger, R., Reiffert, S., Rothe, C., Tesar, M., Thomassen, E., Cao, M., Dreier, T., Fischer, D., Höß, A., Landon, I., Knappik, A., Marget, M., Pack., P., Meng., X.-Q., Schier, R., Söhlemann, P., Winter, J., Wölle, J., Kretzschmar, T. (2001). High-throughput generation and engineering of recombinant human antibodies. Journal of Immunological Methods, 254, 67–84.CrossRefGoogle ScholarPubMed
Löhning, C. (2001). Novel methods for displaying (poly)peptides/proteins on bacteriophage particles via disulfide bonds. WO 01/05950.
Nagy, Z.A., Hubner, B., Löhning, C., Rauchenberger, R., Reiffert, S., Thomassen-Wolf, E., Zahn, S., Leyer, S., Schier, E.M., Zahradnik, A., Brunner, C., Lobenwein, K., Rattel, B., Stanglmaier, M., Hallek, M., Wing, M., Anderson, S., Dunn, M., Kretzschmar, T., Tesar, M. (2002). Fully human, HLA-DR-specific monoclonal antibodies efficiently induce programmed death of malignant lymphoid cells. Nature Medicine, 8, 801–7.CrossRefGoogle ScholarPubMed
Ohara, R., Knappik, A., Shimada, K., Frisch, C., Ylera, F., Koga, H. (2006). Antibodies for proteomic research: comparison of traditional immunization with recombinant antibody technology. Proteomics, 6, 2638–46.CrossRefGoogle ScholarPubMed
Pack, P. and Plückthun, A. (1992). Miniantibodies: use of amphipathic helices to produce functional, flexibly linked dimeric FV fragments with high avidity in Escherichia coli. Biochemistry, 31, 1579–84.CrossRefGoogle ScholarPubMed
Rauchenberger, R., Borges, E., Thomassen-Wolf, E., Rom, E., Adar, R., Yaniv, Y., Malka, M., Chumakov, I., Kotzer, S., Resnitzky, D., Knappik, A., Reiffert, S., Prassler, J., Jury, K., Waldherr, D., Bauer, S., Kretzschmar, T., Yayon, A., Rothe, C. (2003). Human combinatorial Fab library yielding specific and functional antibodies against the human fibroblast growth factor receptor 3. Journal of Biological Chemistry, 278, 38194–205.CrossRefGoogle ScholarPubMed
Rothe, C., Urlinger, S., Löhning, C., Prassler, J., Stark, Y., Jäger, U., Hubner, B., Bardroff, M., Pradel, I., Boss, M., Bittlingmaier, R., Bataa, T., Frisch, C., Brocks, B., Honegger, A., Urban, M. (2008). The Human Combinatorial Antibody Library HuCAL GOLD combines diversification of all six CDRs according to the natural immune system with a novel display method for efficient selection of high-affinity antibodies. Journal of Molecular Biology, 376, 1182–200.CrossRefGoogle ScholarPubMed
Sergeeva, A., Kolonin, M.G., Molldrem, J.J., Pasqualini, R., Arap, W. (2006). Display technologies: application for the discovery of drug and gene delivery agents. Advanced drug Delivery Reviews, 58, 1622–54.CrossRefGoogle ScholarPubMed
Steidl, S., Ratsch, O., Brocks, B., Dürr, M., Thomassen-Wolf, E. (in press). In vitro affinity maturation of human GM-CSF antibodies by targeted CDR-diversification. Molecular Immunology.
Sun, C., Kilburn, D., Lukashin, A., Crowell, T., Gardner, H., Brundiers, R., Diefenbach, B., Carulli, J.P. (2003). Kirrel2, a novel immunoglobulin superfamily gene expressed primarily in β cells of the pancreatic islets. Genomics, 82, 130–42.CrossRefGoogle ScholarPubMed
Virnekäs, B., Ge, L., Plückthun, A., Schneider, K.C., Wellnhofer, G.Moroney, S.E. (1994). Trinucleotide phosphoramidites: ideal reagents for the synthesis of mixed oligonucleotides for random mutagenesis. Nucleic Acids Research, 22, 5600–7.CrossRefGoogle ScholarPubMed
Yang, W.P., Green, K., Pinz-Sweeney, S., Briones, A.T., Burton, D.R., Barbas, C.F.. (1995). CDR walking mutagenesis for the affinity maturation of a potent human anti-HIV-1 antibody into the picomolar range. Journal of Molecular Biology, 254, 392–403.CrossRefGoogle ScholarPubMed

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