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Expression of a chimeric antibody in the baculovirus system

Authors Tan, Wei Qi
Issue Date 1996
Summary The development of hybridoma technology has resulted in an enormous expansion of the use of antibodies in medicine and biology. This technology allows the production of antibodies of defined specificity. Recently, developments in genetic engineering have facilitated the isolation of diverse repertoires of antibody genes from antibody-producing cells followed by their expression in a protein expression system. This technique thus could provide an alternative to hybridoma technology for the production of monoclonal antibodies. In this work, we have expressed a mouse/human chimeric antibody and its Fab fragment against bacterial endotoxin in the baculovirus expression system, which may allow the mass production of secretory recombinant antibodies. In order to target the insect-produced antibody into the culture medium, we have constructed a new dual expression baculovirus transfer vector pPLSP2 by inserting the melittin signal peptide downstream of the polyhedrin promoter and the B.mori larval serum protein signal peptide downstream of the p10 promoter. Our results showed that both signal peptides could target foreign proteins into the culture media. Furthermore, the insect cell could correctly assemble the heavy and the light chains into heterodimeric molecules. The finding that the chimeric antibody could bind to protein A suggested that the antibody was in the correct H2L2 form. Moreover, only the heavy chain in combination with the light chain could bind to antigen LPS, while the light or the heavy chains alone could not. In conclusion, this dual secretary protein expression vector pPLSP2 has proved useful in the expression of active immunoglobulins and probably other heterodimeric proteins in the baculovirus system
Note Thesis (M.Phil.)--Hong Kong University of Science and Technology, 1996
Language English
Format Thesis
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