By Henry Y. Wang and Tadayuki Imanaka (Eds.)
content material: creation of engineered antibodies in myeloma and hybridoma cells : improvements in gene expression and approach layout / D.K. Robinson, D. DiStefano, S.L. Gould, G. Cuca, T.C. Seamans, D. Benincasa, S. Munshi, C.P. Chan, J. Stafford-Hollis, G.F. Hollis, D. Jain, ok. Ramasubramanyan, G.E. Mark, and M. Siberklang --
Antibody construction in human hybridomas with growth-associated creation kinetics / Hidekazu Sawada and Kazuaki Kitano --
Antibody construction in chinese language hamster ovary cells utilizing an impaired selectable marker / R.S. Barnett, K.L. Limoli, T.B. Huynh, E.A. Ople, and M.E. Reff --
Antibody construction in insect cells / Kathleen N. Potter, Yucheng Li, and J. Donald Capra --
Antibody expression in crops / J. K-C. Ma --
Engineering of immunoglobulin Fc and single-chain Fv proteins in Escherichia coli / David Filpula, Michele Rollence, Nina Essig, James Nagle, Aniruddha Achari, and Timothy Lee --
T-cell distinct immunofusion proteins from Escherichia coli : intent, layout, and homes / Marc higher and Patricia A. Nolan --
creation and purification of antibody utilizing Bacillus subtilis as an expression host / S.L. Wong, X.C. Wu, L.P. Yang, S.C. Ng, and N. Hudson --
Trichoderma reesei, a promising novel host for antibody construction / Eini Nyyssönen and Sirkka Keränen --
utilising antibody catalysis to natural synthesis / Brian J. Lavey and Kim D. Janda --
powerful and particular peroxidase job with an antibody L chain-porphyrin Fe(III) advanced / Tadayuki Imanaka and Masahiro Takagi.
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Content material: construction of engineered antibodies in myeloma and hybridoma cells : improvements in gene expression and procedure layout / D. ok. Robinson, D. DiStefano, S. L. Gould, G. Cuca, T. C. Seamans, D. Benincasa, S. Munshi, C. P. Chan, J. Stafford-Hollis, G. F. Hollis, D. Jain, okay. Ramasubramanyan, G.
Extra info for Antibody Expression and Engineering
A higher percentage of the clones surviving selection are those in which the impaired NEO gene has been integrated into 'hot spots' within the genome, which concomitandy yield very high levels of linked gene expression. Once isolated, transfectants which display a very high level of immunoglobulin protein production are induced to undergo gene amplification by selection in Methotrexate (MTX) for the dihydrofolate reductase (DHFR) gene (9). As the DHFR gene copy number increases through amplification, there is a parallel increase in the closely linked immunoglobulin gene copy number with an accompanying rise in immunoglobulin production.
H. (1984) Cell culture methods for molecular and cell biology. Vol. 1-4, Alan R. , New York. 24. , and Kitano, K. (1988) Polyethylene glycols for promoting the growth of mammalian cells. Appl. Microbiol. , 27, 533537. 25. , Kano-Sueoka, T. (1982) Growth of hybridoma cells in serum-free medium: Ethanolamine is an essential component. Proc. Natl. Acad. , 79, 1158-1162. 26. , and Kida, M. (1986). Production of human monoclonal antibodies by heterohybridomas. Appl. Microbiol. Biotechnol 24, 282-286.
The immunoglobulin domain structure can be manipulated by exchanging whole domains or portions of domains between antibodies. Synthetic antibodies have been produced in many expression systems including bacteria, yeast, plant, mammalian and insect cells (reviewed in 3). Some of the difficulties involved in studying human antibodies are the result of technical problems such as obtaining stable human-human hybridomas and the lack of secreted monoclonal antibodies in amounts sufficient for analysis.