Conjugation of metallothionein to a murine monoclonal antibody.
A method of conjugation of the metal-binding protein, metallothionein, to an anticarcinoma murine monoclonal antibody, B72.3, and its F(ab')2 fragment has been developed utilizing the heterobifunctional crosslinking reagent, succinimidyl 4-(N-maleimidomethyl)-cyclohexane 1-carboxylate. This crosslinking reagent is first reacted with the free amines on the immunoglobulin. After removal of unreacted crosslinker, conjugation is affected through a sulfhydryl group on metallothionein. Under the conditions employed all immunoglobulin aggregates contained metallothionein. The degree of undesired aggregation is directly proportional to the number of metallothioneins attached to the immunoglobulin. This aggregation can be controlled by the amount of crosslinker and metallothionein presented to the immunoglobulin. The immunoglobulin conjugate retains full immunoreactivity and can be readily purified from the unreacted metallothionein and high molecular weight aggregates. The metallothionein-B72.3 conjugate functions as an efficient and stable chelator of radiometals. Thus metallothionein-monoclonal antibody conjugates have potential utility in cancer diagnosis and therapy.
Brown
B A
BA
Medical Products Department, E.I. DuPont & Company, N. Billerica, Massachusetts 01862.
Drozynski
C A
CA
Dearborn
C B
CB
Hadjian
R A
RA
Liberatore
F A
FA
Tulip
T H
TH
Tolman
G L
GL
Haber
S B
SB
D000911:Antibodies, Monoclonal; D055598:Chemical Phenomena; D002621:Chemistry; D002845:Chromatography / Q000379:methods; D003432:Cross-Linking Reagents; D007553:Isotope Labeling; D008301:Maleimides; D008668:Metallothionein; D013667:Technetium
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D008301:Maleimides; C025285:N-(4-carboxycyclohexylmethyl)maleimide N-hydroxysuccinimide ester; D013667:Technetium; D008668:Metallothionein