SpringerProtocols: Abstract: 13. Production of Recombinant Bispecific Antibodies

13. Production of Recombinant Bispecific Antibodies

By: Roland E. Kontermann², Tina Völkel², Tina Korn³

Abstract

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Recombinant bispecific antibodies have great potential as diagnostic and therapeutic reagents (1,2). Applications of recombinant bispecific antibodies include the recruitment of effector molecules (e.g., enzymes, complement components, immunoglobulins), effector cells (cytotoxic T-lymphocytes, natural killer [NK] cells, macrophages), or gene-therapeutic vectors (e.g., adenoviruses) to other molecules or target cells (3–10). Through genetic engineering, molecules with the desired properties can be generated and produced in various prokaryotic or eukaryotic expression systems (11). For example, by starting from antibody fragments isolated from human combinatorial libraries by phage display, completely human bispecific molecules can be generated.

Affiliation(s): (2) Vectron Therapeutics AG, Marburg, Germany
(3) Institut für Molekularbiologie und Tumorforschung, Marburg, Germany

Book Title: Antibody Engineering: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 248 | Pub. Date: Dec-05-2003 | Page Range: 227-242 | DOI: 10.1385/1-59259-666-5:227

Subject: Biochemistry

References

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1.	Segal, D. M., Weiner, G. J., and Weiner, L. M. (1999) Bispecific antibodies in cancer therapy. Curr. Opin. Immunol. 11, 558–562.

2.	Withoff, S., Helfrich, W., de Leij, L. F., and Molema, G. (2001) Bispecific antibody therapy for the treatment of cancer. Curr. Opin. Mol. Ther. 3, 353–362.

3.	Holliger, P., Prospero, T. D., and Winter, G. (1993) “Diabodies”, small bivalent and bispecific antibody fragments. Proc. Natl. Acad. Sci. USA 90, 6444–6448.

4.	Kontermann, R. E., Martineau, P., Cummings, C. E., Karpas, A., Allen, D., Derbyshire, E., et al. (1997) Enzyme immunoasssays using bispecific diabodies. Immunotechnology 3, 137–144.

5.	Kontermann, R. E., Wing, M. G., and Winter, G. (1997) Complement recruitment using bispecific diabodies. Nat. Biotechnol. 15, 629–631.

6.	Löffler, A., Kufer, P., Lutterbuse, R., Zettl, F., Daniel, P. T., Schwenkenbecker, J. M., et al. (2000) A recombinant bispecific single-chain antibody, CD19×CD2, induces rapid and high lymphoma-directed cytotoxicity by unstimulated T lymphocytes. Blood 95, 2098–2103.

7.	Brüsselbach, S., Korn, T., Völkel, T., Müller, R., and Kontermann, R. E. (1999) Enzyme recruitment and tumor cell killing in vitro by a bispecific single-chain diabody. Tumor Targeting 4, 115–123.

8.	Cochlovius, B., Kipriyanov, S. M., Stassar, M. J., Schuhmacher, J., Benner, A., Moldenhauer, G., et al. (2000) Cure of Burkitt’s lymphoma in severe combined immunodeficiency mice by T cells, tetravalent CD3×CD19 tandem diabody, and CD28 costimulation. Cancer Res. 60, 4336–4341.

9.	Haisma, H. J., Grill, J., Curiel, D. T., Hoogeland, S., van Beusechem, V. W., Pinedo, H. M., et al. (2000) Targeting of adenoviral vectors through a bispecific single-chain antibody. Cancer Gene Ther. 7, 901–904.

10.	Nettelbeck, D. M., Miller, D. W., Jerome, J., Zuzarte, M., Watkins, S. J., Hawkins, R. E., et al. (2001) Retargeting of adenovirus to endothelial cells by a bispecific single-chain diabody directed against the adenovirus knob domain and human endoglin (CD105). Mol. Ther. 3, 882–891.

11.	Kontermann, R. E. and Dübel, S. (2001) Antibody Engineering. Lab Manual Series-Springer-Verlag, Heidelberg.

12.	Kriangkum, J., Xu, B., Nagata, L. P., Fulton, R. E., and Suresh, M. R. (2001) Bispecific and bifunctional single chain recombinant antibodies. Biomolecular Eng. 18, 31–40.

13.	Kipriyanov, S. M., Moldenhauer, G., Schuhmacher, J., Cochlovius, B., von der Lieth, C. W., Matys, E. R., et al. (1999) Bispecific tandem diabody for tumor therapy with improved antigen binding and pharacokinetics. J. Mol. Biol. 293, 41–56.

14.	Alt, M., Müller, R., and Kontermann, R. E. (1999) Novel tetravalent and bispecific IgG-like antibody molecules combining single-chain diabodies with the immunoglobulin γ1 Fc or CH3 region. FEBS Lett. 454, 90–94.

15.	Park, S. S., Ryu, C. J., Kang, Y. J., Kashmiri, S.V.S., and Hong, H. J. (2000) Generation and characterization of a novel tetravalent bispecific antibody that binds to hepatitis B virus surface antigens. Mol. Immunol. 37, 1123–1130.

16.	Kontermann, R. E. and Müller, R. (1999) Intracellular and cell surface displayed single-chain diabodies. J. Immunol. Methods 226, 179–188.

17.	Völkel, T., Korn, T., Bach, M., Müller, R., and Kontermann, R. E. (2001) Optimized linker sequences for the expression of monomeric and dimeric single-chain diabodies. Protein Eng. 14, 815–823.

18.	Mallender, W. D., Ferreira, S. T., Voss, E. W. Jr., and Coelho-Sampaio, T. (1994) Inter-active-site distance and solution dynamics of a bivalent-bispecific single-chain antibody molecule. Biochemistry 33, 10100–10108.

19.	Gruber, M., Schodin, B. A., Wilson, E. R., and Kranz, D. M. (1994) Efficient tumor cell lysis mediated by a bispecific single chain antibody expressed in Escherichia coli. J. Immunol. 152, 5368–5374.

20.	Hayden, M. S., Linsley, P. S., Gayle, M. A., Bajorath, J., Brady, W. A., Norris, N. A., et al. (1994) Single-chain mono-and bispecific antibody derivatives with novel biological properties and antitumor activity from a COS cell transcient expression system. Ther. Immunol. 1, 3–15.

21.	Fischer, R., Schumann, D., Zimmermann, S., Drosard, J., Sack, M., and Schillberg, S. (1999) Expression and characterization of bispecific single-chain Fv fragments produced in transgenic plants. Eur. J. Biochem. 262, 810–816.

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