Production of a monoclonal antibody against the thrombin-like enzyme, habutobin, from Trimeresurus flavoviridis venom.
We succeeded in producing a monoclonal antibody to the thrombin-like enzyme, habutobin, which was purified from crude venom of the snake Trimeresurus flavoviridis. The monoclonal antibody obtained belonged to IgG1, and its light chain consisted of a kappa-chain. The monoclonal antibody reacted specifically with habutobin and crude venom from T. flavoviridis but did not react with human thrombin or bovine thrombin on Western blotting. The concentration of habutobin and crude venom of T. flavoviridis, in vitro, could be measured by means of ELISA using the monoclonal antibody. Furthermore, the ELISA-double sandwich method employing this monoclonal antibody may represent a reliable method for determining the habutobin levels in the circulating blood.
Nakamura
M
M
Department of Physiology, School of Medicine, University of the Ryukus, Okinawa, Japan.
Kinjoh
K
K
Kosugi
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D015153:Blotting, Western; D002021:Buffers; D002460:Cell Line; D002846:Chromatography, Affinity; D003435:Crotalid Venoms / Q000201:enzymology* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D007074:Immunoglobulin G / Q000276:immunology; D007202:Indicators and Reagents; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012697:Serine Endopeptidases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D002021:Buffers; D003435:Crotalid Venoms; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D012697:Serine Endopeptidases; C078095:habutobin