TY - JOUR
T1 - Production of genetically engineered antibodies in myeloma cells
T2 - Design, expression, and applications
AU - Wright, Ann
AU - Shin, Seung Uon
N1 - Funding Information:
We thank Sherie L. Morrison for her support and helpful comments on the manuscript. This work was supported in part by Grants (to S.L.M.) CA 16858, AI 19041, and AI 129470 from the National Institutes of Health, and by Grant IM-603 from the American Cancer Society.
PY - 1991/4
Y1 - 1991/4
N2 - The criteria for producing recombinant antibodies in myeloma cells are discussed. Heavy- and light-chain genes are cloned into expression vectors with different selectable markers and transfected simultaneously into recipient cells. Expression vectors that contain not only the elements required for gene expression but features that facilitate the exchange of variable or constant region genes have been designed. Obtaining variable region genes of interest has been greatly facilitated by the advent of the polymerase chain reaction. Murine variable region genes may be joined to human constant regions, whose domain structure has been exploited by the design of cloning "cassettes." In these, unique restriction sites placed between constant region exons allow for domain duplication, deletion, or exchange. Immunoglobulin fusion proteins, in which antigen specificities have been fused to heterologous proteins, have been produced successfully. Certain antibody properties have also been altered by site-directed mutagenesis. These manipulations not only have provided useful insights into structure-function relationships in antibodies, but enable the production of antibodies with altered or novel properties.
AB - The criteria for producing recombinant antibodies in myeloma cells are discussed. Heavy- and light-chain genes are cloned into expression vectors with different selectable markers and transfected simultaneously into recipient cells. Expression vectors that contain not only the elements required for gene expression but features that facilitate the exchange of variable or constant region genes have been designed. Obtaining variable region genes of interest has been greatly facilitated by the advent of the polymerase chain reaction. Murine variable region genes may be joined to human constant regions, whose domain structure has been exploited by the design of cloning "cassettes." In these, unique restriction sites placed between constant region exons allow for domain duplication, deletion, or exchange. Immunoglobulin fusion proteins, in which antigen specificities have been fused to heterologous proteins, have been produced successfully. Certain antibody properties have also been altered by site-directed mutagenesis. These manipulations not only have provided useful insights into structure-function relationships in antibodies, but enable the production of antibodies with altered or novel properties.
UR - http://www.scopus.com/inward/record.url?scp=1842397697&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=1842397697&partnerID=8YFLogxK
U2 - 10.1016/S1046-2023(05)80213-0
DO - 10.1016/S1046-2023(05)80213-0
M3 - Article
AN - SCOPUS:1842397697
VL - 2
SP - 125
EP - 135
JO - Methods in Enzymology
JF - Methods in Enzymology
SN - 0076-6879
IS - 2
ER -