Development of a monoclonal antibody against ochratoxin A and its application in enzyme-linked immunosorbent assay and gold nanoparticle immunochromatographic strip.
A monoclonal antibody (mAb) specific to ochratoxin A (OTA) was produced from a stable hybridoma cell line, 9C9H9, generated by the fusion of P3/NS1/1-AG4-1 myeloma cells with spleen cells isolated from a BALB/c mouse immunized with OTA-keyhole limpet hemocyanin. The 9C9H9 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. The concentrations causing 50% inhibition of binding of OTA-horseradish peroxidase to the antibody by OTA, OTB, and OTC were found to be 0.32, 0.17, and 0.28 ng/mL, respectively, in the cdELISA. A sensitive and rapid mAb-based gold nanoparticle immunochromatographic strip was also developed using this mAb. This strip has a detection limit of 5 ng/mL for OTA and can be completed in 10 min. Analysis of OTA in coffee samples revealed that data obtained from immunochromatographic strip were in a good agreement with those obtained from cdELISA. The mAb-based cdELISA and immunochromatographic strip assay established in this study were sensitive and accurate for rapid screening of OTA in coffee samples.
Liu
Biing-Hui
BH
Department of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan.
Tsao
Zih-Jay
ZJ
Wang
Jing-Jhih
JJ
Yu
Feng-Yih
FY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D002417:Cattle; D003069:Coffee / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005506:Food Contamination / Q000032:analysis; D006046:Gold / Q000737:chemistry / Q000378:metabolism*; D053768:Metal Nanoparticles / Q000737:chemistry*; D051379:Mice; D009793:Ochratoxins / Q000032:analysis* / Q000276:immunology; D011934:Reagent Strips; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D003069:Coffee; D009793:Ochratoxins; D011934:Reagent Strips; C025589:ochratoxin A; D006046:Gold