Antibody Expression and Engineering by Henry Y. Wang and Tadayuki Imanaka (Eds.)

By Henry Y. Wang and Tadayuki Imanaka (Eds.)

content material: construction of engineered antibodies in myeloma and hybridoma cells : improvements in gene expression and method 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 creation 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 detailed immunofusion proteins from Escherichia coli : reason, layout, and homes / Marc greater 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 --
employing antibody catalysis to natural synthesis / Brian J. Lavey and Kim D. Janda --
powerful and particular peroxidase task with an antibody L chain-porphyrin Fe(III) advanced / Tadayuki Imanaka and Masahiro Takagi.

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Colonies were assayed for expression by taking supernatant samples from confluent wells. Due to variations in cell number when assayed, antibody present in the well from residual media, and the experimental error of the ELIS A assay itself, the absolute expression values obtained are not precise. However, the wide range of immunoglobulin expression level observed in this experiment, from less than 25ng/ml to greater than 500 ng/ml, can not be explained by solely by sampling variables. When expanded and more accurate expression values are obtained, clones still show large differences in expression levels.

BARNETT ET AL. ; ACS Symposium Series; American Chemical Society: Washington, DC, 1995. ch003 30 ANTIBODY EXPRESSION AND ENGINEERING chimeric mouse-human anti-CD20 antibody was transfected into Chinese Hamster Ovary (CHO) cells. Two hundred fifty eight (258) individual G418 resistant colonies were assayed for immunoglobulin production in 96 well dishes. Colonies were assayed for expression by taking supernatant samples from confluent wells. Due to variations in cell number when assayed, antibody present in the well from residual media, and the experimental error of the ELIS A assay itself, the absolute expression values obtained are not precise.

Press, Tokyo and Springer Verlag, Berlin. 19. , and Kitano, K. (1990) Improvement in the antibody productivity of humna-human hybridomas by transfection with Tac gene. Cytotechnology, 4, 29-37. 20. , and Kitano, K. (1991) Improvement in the proliferative activity of human-human hybridomas at low cell density by transfection with bFGF gene. Cytotechnology, 6, 90-103. 21. , and Akiyama, S. (1988) Effective production of a human monoclonal antibody against tetanus toxoid by selection of high productivity clones of a heterohybridoma.

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