By Henry Y. Wang and Tadayuki Imanaka (Eds.)
content material: construction 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 construction kinetics / Hidekazu Sawada and Kazuaki Kitano --
Antibody creation 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 specific immunofusion proteins from Escherichia coli : motive, layout, and houses / Marc higher and Patricia A. Nolan --
construction 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 --
robust and particular peroxidase job with an antibody L chain-porphyrin Fe(III) complicated / Tadayuki Imanaka and Masahiro Takagi.
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Extra info for Antibody Expression and Engineering
1%) expressed less than 25 ng/ml of antibody. 0 χ 10 cells/electroporation) of the TCAE vector, which contains a consensus Kozak sequence at the translation initiation site, yielded 258 G418 resistant colonies. There were no colonies in this experiment that produced greater than 1000 ng/ml of antibody. Two hundred and one (201) of these colonies (78%) expressed less than 25 ng/ml of antibody. In other experiments where equal amounts of DNA of the two vectors (TCAE, ANEX) were transfected simultaneously, the ANEX vector consistendy yielded a 80-90% reduction in numbers of G418 resistant clones per electroporation.
As a result of comparison of various culture methods, we decided that short-term perfusion culture should be a practical method for h-h hybridomas with growth-associated production kinetics. 29AGC-1 was performed under short-term perfusion using a 50-L fermentor, and the purification of IgG-MoAb was efficiendy carried out in yields of approximately 75% on the scale of more than 100L of culture supernatant. These IgG MoAbs neutralized HBV infectivity and protected chimpanzees against the development of hepatitis B.
BARNETT ET AL. ch003 1 2 3 4 Figure 8 Shown is a Northern blot of RNA isolated from a G418 resistant TCAE clone and NEOSPLA clone. Both clones secreted high amounts of chimeric antibody. ; ACS Symposium Series; American Chemical Society: Washington, DC, 1995. ch003 1 2 3 4 5 Figure 9 Shown is a Southern Blot of DNA isolated from a CHO clone integrated into a 'hot spot' and the derived 5nM and 50nM MTX clones. 5 mg of high molecular weight DNA was digested with Nhe I which cuts once in each plasmid and gives two bands of unknown size for each integration site into cellular DNA.
Antibody Expression and Engineering by Henry Y. Wang and Tadayuki Imanaka (Eds.)