Monoclonal antibody-based lateral flow assay for detection of botulinum neurotoxin type A.
A rapid lateral flow assay was developed to detect botulinum neurotoxin type A (BoNT/A). The assay was based on the sandwich format using monoclonal antibodies (MAbs) of two distinct specificities. One anti-BoNT/A heavy chain MAb (150-3) was immobilized to a defined detection zone on a porous nitrocellulose membrane, while the other anti-BoNT/A heavy chain MAb (44.1) was conjugated to colloidal gold particles, which served as a detection reagent. The BoNT/A-containing sample was added to the strip and allowed to react with MAb (44.1)-coated particles. The mixture was then passed along the porous membrane by capillary action past the MAb (150-3) in the detection zone, which binds the particles that had BoNT/A bound to their surface, giving a red color within this detection zone with intensity proportional to BoNT/A concentration. In the absence of BoNT/A, no immunogold was bound to the solid-phase antibody. With this method, 50 ng/mL of BoNT/A were detected in less than 10 min. The assay sensitivity can be increased by silver enhancement to 1 ng/mL. The developed BoNT/A assay also showed no cross-reaction to type B neurotoxin (BoNT/B) and type E neurotoxin (BoNT/E).
Chiao
Der-Jiang
DJ
Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan, Republic of China.
Wey
Jiunn-Jye
JJ
Shyu
Rong-Hwa
RH
Tang
Shiao-Shek
SS
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000937:Antigen-Antibody Reactions; D019274:Botulinum Toxins, Type A / Q000032:analysis*; D017979:Gold Colloid / Q000737:chemistry; D007118:Immunoassay / Q000379:methods
D000911:Antibodies, Monoclonal; D017979:Gold Colloid; D019274:Botulinum Toxins, Type A