Assembly of functional antibodies from immunoglobulin heavy and light chains synthesised in E. coli.
Genes for a murine mu heavy chain and a lambda light chain immunoglobulin have been inserted into bacterial expression plasmids containing the Escherichia coli trp promoter and ribosome binding site. Induction of transcription from the trp promoter results in accumulation of both light and heavy chain polypeptides in appropriate host strains. Both proteins were found as insoluble products. Following extraction and purification of the immunoglobulin containing fractions, antigen binding activity was recovered. The activity demonstrates essentially the same properties as the antibody from the hybridoma from which the genes were cloned.
D000818:Animals; D000906:Antibodies / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D006241:Haptens; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007146:Immunoglobulin lambda-Chains / Q000235:genetics / Q000302:isolation & purification; D007148:Immunoglobulin mu-Chains / Q000235:genetics / Q000302:isolation & purification; D007700:Kinetics; D051379:Mice; D010957:Plasmids
D000906:Antibodies; D006241:Haptens; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007146:Immunoglobulin lambda-Chains; D007148:Immunoglobulin mu-Chains; D004262:DNA Restriction Enzymes