Development of a monoclonal antibody against domoic acid and its application in enzyme-linked immunosorbent assay and colloidal gold immunostrip.
A monoclonal antibody (mAb) specific to domoic acid was produced from a stable hybridoma cell line, 9F1F11, generated by the fusion of P3/NS1/1-AG4-1 myeloma cells with spleen cells isolated from a Balb/c mouse immunized with domoic acid--keyhole limpet hemocyanin. The 9F1F11 mAb belongs to the immunoglobulin G1 (kappa-chain) isotype. A competitive direct enzyme-linked immunosorbent assay (cdELISA) and a competitive indirect ELISA were established for antibody characterization. In the cdELISA, the concentration causing 50% inhibition (IC50) of binding of domoic acid-horseradish peroxidase to the antibody by domoic acid was found to be 0.58 ng/mL. A sensitive and rapid mAb-based colloidal gold immunostrip was also developed. The immunostrip assay, which has a detection limit of 5 ng/mL for domoic acid, can be completed in 10 min. Analysis of domoic acid in blue mussel samples revealed that data obtained from immunostrip were in a good agreement with those obtained from cdELISA. The mAb-based cdELISA and immunostrip assay established in this study were sensitive and accurate for rapid screening of domoic acid in shellfish samples.
Tsao
Zih-Jay
ZJ
Department of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan, and Tian-Sheng Memorial Hospital, Tong Kong, Ping-Tong, Taiwan.
Liao
Yi-Chun
YC
Liu
Biing-Hui
BH
Su
Ching-Chyuan
CC
Yu
Feng-Yih
FY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D017979:Gold Colloid; D007608:Kainic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D008387:Marine Toxins / Q000032:analysis; D049880:Mytilus edulis / Q000737:chemistry; D009467:Neuromuscular Depolarizing Agents / Q000032:analysis; D009498:Neurotoxins / Q000032:analysis; D011934:Reagent Strips; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000032:analysis
D000911:Antibodies, Monoclonal; D017979:Gold Colloid; D008387:Marine Toxins; D009467:Neuromuscular Depolarizing Agents; D009498:Neurotoxins; D011934:Reagent Strips; C012301:domoic acid; D007608:Kainic Acid