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 strategy 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 construction 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 certain immunofusion proteins from Escherichia coli : reason, layout, and homes / Marc greater 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 --
employing antibody catalysis to natural synthesis / Brian J. Lavey and Kim D. Janda --
robust and particular peroxidase task with an antibody L chain-porphyrin Fe(III) advanced / Tadayuki Imanaka and Masahiro Takagi.

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The process of transfection usually involves the simultaneous introduction of the transfer vector containing the gene of interest and wild-type virus DNA into the same cell. Homologous recombination between the vector and the wild-type virus DNA results in the viral genome containing the foreign gene. There are several ways to introduce DNA into insect cells. Transfection of insect cells is often performed using calcium phosphate precipitation (36,37,38), and electroporation has been reported as an efficient method (39).

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. ; ACS Symposium Series; American Chemical Society: Washington, DC, 1995. ch003 3. BARNETT ET AL. CHO CeUs Using an Impaired Selectable Marker 39 were screened, and the highest immunoglobulin expresser was expanded.

Some transfer vectors are designed for the coexpression of two, three and four different proteins (reviewed in 12 and 32, 33). There are vectors which allow for a color distinction between wild-type and recombinant virus plaques, thereby reducing the effort required to plaque purify recombinant viruses. Some vectors also contain sequences which result in the production of recombinant proteins fused to tags which are useful in the rapid purification of the recombinant protein from insect cellular and viral proteins.

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