Monoclonal antibody-based enzyme immunoassay for detection of botulinum neurotoxin type A.
A sensitive and specific ELISA was developed to detect BoNT/A in biological fluids. The assay is based on the sandwich format using monoclonal antibodies (MAb) of two distinct specificities. An affinity-purified anti-BoNT/A heavy chain MAb (150-3) is utilized to adsorb BoNT/A from solution; the second anti-BoNT/A heavy chain MAb (44-1A) conjugated with peroxidase is then used to form a sandwich. Peroxidase allows color development and measurement of optical density at 450 nm. Standard curves were linear over the range of 2.5 to 100 ng/mL BoNT/A. The limit of detection was below 5 ng/mL in assay buffer, as well as in a 1:10 dilution of urine or 1:50 dilution of human serum spiked with BoNT/A. 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-Jey
JJ
Tang
Shiao-Shek
SS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D019274:Botulinum Toxins, Type A / Q000032:analysis* / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000302:isolation & purification; D008163:Luminescent Measurements; D051379:Mice; D008807:Mice, Inbred BALB C; D009195:Peroxidase / Q000737:chemistry
D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D009195:Peroxidase; D019274:Botulinum Toxins, Type A