Purification of human glutamate dehydrogenase (GDH) and an adsorptive voltammetric investigation of the interaction of GDH with rabbit anti-human GDH antibody.
A procedure for the isolation of glutamate dehydrogenase (GDH) from human liver, which involves the use of ion-exchange chromatography on diethylaminoethyl cellulose and affinity chromatography on guanosine triphosphate conjugated to Sepharose 4B, is described. The adsorptive voltammetric behaviour of human GDH, bovine GDH and rabbit anti-human GDH antibody was optimised with respect to accumulation potential, accumulation time and scan rate. The lower limits of detection were 0.2 and 1.2 mg l-1 for human and bovine GDH, respectively, and the lower limit of detection for rabbit anti-GDH antibody was 0.04 mg l-1. The interaction of human GDH with rabbit anti-human GDH antibody was also examined using this method.
Carty
P
P
School of Biological Sciences, Dublin City University, Ireland.
O'Kennedy
R
R
Lorenzo Abad
E
E
Fernández Alvarez
J M
JM
Rodriguez Flores
J
J
Smyth
M R
MR
Tipton
K
K
D000818:Animals; D000937:Antigen-Antibody Reactions; D002417:Cattle; D005969:Glutamate Dehydrogenase / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D008099:Liver / Q000201:enzymology; D011817:Rabbits / Q000276:immunology
D005969:Glutamate Dehydrogenase