An ISFET-based immunosensor for the detection of beta-Bungarotoxin.
An ion-sensitive field effect transistor (ISFET)-based immunosensor was developed to detect/quantitate beta-Bungarotoxin (beta-BuTx), a potent presynaptic neurotoxin from the venom of Bungarus multicinctus. A murine monoclonal antibody (mAb 15) specific to beta-BuTx was immobilized onto silicon nitride wafers after silanization and activation with glutaraldehyde. A chip based enzyme linked-immunosorbantassay (ELISA) was performed to ascertain antigen binding to the immobilized antibody. To develop an electrochemical immunosensing system for the detection/quantitation of beta-BuTx, an ISFET was used as a solid phase detector. MAb 15 was immobilized on the gate region of the ISFET. The antigen antibody reaction was monitored by the addition of urease conjugated rabbit anti-beta-BuTx antibodies. The sensor can detect toxin level as low as 15.6 ng/ml. The efficacy of the sensor for the determination of beta-BuTx from B. multicinctus venom was demonstrated in mouse model. Toxin concentration was highest at the site of injection (748.0+/-26 ng/ml) and moderate amount was found in the plasma (158.5+/-13 ng/ml).
Department of Anatomy, Venom and Toxin Research Programme, Faculty of Medicine, National University of Singapore, 4 Medical Drive, Singapore, 117597, Singapore. selvanze@umdnj.edu
Z Emmanuel
ZE
Selvanayagam
P
P
Neuzil
P
P
Gopalakrishnakone
U
U
Sridhar
M
M
Singh
L C
LC
Ho
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002038:Bungarotoxins / Q000032:analysis* / Q000097:blood / Q000276:immunology; D017838:Bungarus; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D007477:Ions; D008297:Male; D051379:Mice; D011817:Rabbits; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D040521:Subcutaneous Tissue / Q000737:chemistry / Q000276:immunology; D014173:Transistors, Electronic
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002038:Bungarotoxins; D007477:Ions