Plastic enzyme-linked immunosorbent assays (ELISA)-on-a-chip biosensor for botulinum neurotoxin A.
A plastic ELISA-on-a-chip (EOC) employing the concept of cross-flow immuno-chromatographic analysis was applied to the measurement of botulinum neurotoxin A (BoNT/A) as agent for bio-terrorism. Two monoclonal antibodies specific to the heavy chain of the toxin were raised and identified to form sandwich binding complexes as the pair with the analyte. For the construction of an immuno-strip, one was utilized as the capture antibody immobilized onto nitrocellulose membrane and the other as the detection coupled to an enzyme, horseradish peroxidase. The two plates of EOC used in this study were fabricated by injection molding of polycarbonate to improve the reproducibility of manufacture and, after inclusion of the immuno-strip, bonded using a UV-sensitive adhesive. Under optimal conditions of analysis, the chip produced a color signal in proportion to the analyte dose and the signal was quantified using a detector equipped with a digital camera. From the dose-response curve, the detection limit of BoNT/A was 2.0 ng mL(-1), approximately five times more sensitive than a commercial-version detection kit employing colloidal gold tracer.
Han
Seung-Mok
SM
Program for Bio-Microsystem Technology, Korea University, 1, 5-ka Anam-dong, Sungbuk-ku, Seoul 136-701, South Korea.
Cho
Joung-Hwan
JH
Cho
Il-Hoon
IH
Paek
Eui-Hwan
EH
Oh
Hee-Bok
HB
Kim
Bong-Su
BS
Ryu
Chunsun
C
Lee
Kyunghee
K
Kim
Young-Kee
YK
Paek
Se-Hwan
SH
D000269:Adhesives; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015374:Biosensing Techniques; D019274:Botulinum Toxins, Type A / Q000032:analysis*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002845:Chromatography / Q000379:methods; D003101:Collodion / Q000737:chemistry; D003124:Colorimetry / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007158:Immunologic Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D011076:Polycarboxylate Cement / Q000737:chemistry; D013154:Spleen / Q000378:metabolism
D000269:Adhesives; D000911:Antibodies, Monoclonal; D011076:Polycarboxylate Cement; C005506:polycarbonate; D003101:Collodion; D019274:Botulinum Toxins, Type A