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PMID: 3508296 Published · ppublish English Journal Article

Expression in COS cells of a mouse-human chimaeric B72.3 antibody.

Protein engineering ·Vol. 1 ·No. 6 ·1987-12-00 ·Pages 499-505

Whittle N, Adair J, Lloyd C, Jenkins L, Devine J, Schlom J, Raubitschek A, Colcher D, Bodmer M

Abstract

B72.3 is a mouse hybridoma cell-line secreting an IgG1 antibody which recognises an epitope on a tumour-associated antigen, TAG-72. This high molecular weight mucin-like molecule is found on a variety of human neoplasms, including colon, breast and ovarian carcinomas. Chimaeric immunoglobulin genes with the B72.3 specificity have been constructed by joining the mouse variable regions from cDNA clones to human genomic constant regions using recombinant DNA techniques. The chimaeric heavy and light chain immunoglobulin genes were placed under the control of a strong viral promoter, and co-transfected into COS-1 cells. SDS-PAGE analysis of the 35S-labelled products demonstrated that the transiently expressed antibodies were correctly synthesised and assembled. The specific binding characteristics of the parent B72.3 antibody were retained by the chimaeric antibody in an antigen-based ELISA. This system gave sufficiently high transient expression of the chimaeric antibody molecules to allow rapid physical and immunological characterisation of the engineered gene products.

MeSH Terms
Amino Acid Sequence Animals Antibodies/genetics Base Sequence Cells, Cultured Chimera Cloning, Molecular DNA/genetics Enzyme-Linked Immunosorbent Assay Genetic Vectors Glycosylation Humans Hybridomas Immunoglobulin G/genetics Mice Molecular Sequence Data Mutation Transfection
Chemicals
Antibodies Immunoglobulin G DNA
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Whittle N
Department of Molecular Immunology, Celltech Limited, Slough, UK.
Adair J
Lloyd C
Jenkins L
Devine J
Schlom J
Raubitschek A
Colcher D
Bodmer M
Article Info
Journal
Protein engineering
Abbr.
Protein Eng
ISSN
0269-2139
Published
1987-12-00
Pages
499-505
Language
English
Region
England
NLM ID
8801484
Subset
IM
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