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Cassette Baculovirus Vectors for the Production of Chimeric, Humanized, or Human-Antibodies in Insect Cells

Poul, M. A.; Cerutti, M.; Chaabihi, H.; Ticchioni, M.; Deramoudt, F. X.; Bernard, A.; Devauchelle, G.; Kaczorek, M.; Lefranc, M. P.

European Journal of Immunology

1995-07 / vol 25 / pages 2005-2009

Abstract

Plasmid cassette-transfer vectors pBHuC kappa, and pBHuC gamma 1 have been designed which enable the construction of recombinant baculoviruses directing the coexpression of complete immunoglobulin in insect cells. We describe the application of these vectors for the expression of a human/mouse chimeric monoclonal antibody of potential immunosuppressive clinical value derived from a mouse anti-human CD29 monoclonal antibody (Mu-K20). The chimeric K20 light and heavy chains produced in sf9 insect cells were correctly processed and assembled into a normal immunoglobulin which is secreted into the culture medium of infected cells. The chimeric mAb Ch-K20-sf9 reproduces in vitro the functional properties of the parental mouse K20: including affinity and inhibition of lymphocyte proliferation. These results demonstrate that the baculovirus/insect cell expression system is suitable for the expression of fully active monoclonal antibodies of therapeutic value. Our generic cassette approach makes this system a very flexible and convenient one for the rapid production of either chimeric, humanized or human mAb with heavy and light chains of any isotype.

0014-2980

Tags

mammalian-cells; secretion; immunoglobulin; baculovirus; t-cells; escherichia-coli; cd29 molecule; chimeric antibody; engineered antibody; expression system; fragment; high-level production; insect cells; integrins; manipulation

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