Purification and characterization of a chimeric bifunctional antibody specific for human carcinoembryonic antigen and indium-benzyl-EDTA.
We describe the purification and characterization of a genetically engineered mouse/human chimeric bifunctional antibody specific for human carcinoembryonic antigen and indium-benzyl-EDTA. A clone expressing the bifunctional antibody has been previously described by our group and was found in this investigation also to express monospecific antibodies as well as Ig forms with mismatched light and heavy chains. The physicochemical properties of these various chimeric immunoglobulins were nearly identical. Isoelectric focusing showed that all these immunoglobulins have pI values between 8.47 and 8.80. A purification method that separates the bifunctional antibody from other Ig forms expressed in the same clone has been devised by relying on a unique interaction between the metal chelate binding region of these antibodies and the sulfopropyl functional group of a TSK SP 5-PW column.
Beidler
D E
DE
Department of Protein Engineering, Hybritech Incorporated, San Diego, California 92196.
Johnson
M J
MJ
Unger
B W
BW
Phelps
J L
JL
Jue
R A
RA
D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000918:Antibody Specificity; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D002678:Chimera; D003432:Cross-Linking Reagents; D004492:Edetic Acid / Q000031:analogs & derivatives* / Q000276:immunology; D006801:Humans; D007204:Indium; D007526:Isoelectric Point; D015202:Protein Engineering
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D003432:Cross-Linking Reagents; D007204:Indium; C069303:benzyl-EDTA; D004492:Edetic Acid