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Development of a new rabbit monoclonal antibody and its based competitive indirect enzyme‐linked immunosorbent assay for rapid detection of sulfonamides
Author(s) -
Liu Na,
Han Zheng,
Lu Lei,
Wang Lin,
Ni Geng,
Zhao Zhihui,
Wu Aibo,
Zheng Xiaodong
Publication year - 2013
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.5945
Subject(s) - monoclonal antibody , sulfapyridine , sulfadiazine , chemistry , chromatography , immunoassay , urine , enzyme , antibody , microbiology and biotechnology , biology , biochemistry , antibiotics , immunology , organic chemistry
Background Monoclonal antibodies generally obtained through the classic mouse hybridoma system were requisite for the establishment of various immunoassays. In this study, a new rabbit monoclonal antibody ( RabMAb ) against sulfonamides ( SAs ) was first produced via hybridoma technique in rabbit. The related enzyme‐linked immunosorbent assay ( ELISA ) was then developed and applied to real sample analysis.Results A sensitive competitive indirect ELISA method based on a novel RabMAb for rapid detection of sulfonamides was first established. The obtained half‐maximum inhibition concentration ( IC 50 ) values for four SAs were all below 10 ng mL −1 , with 0.68 ng mL −1 sulfathiazole ( STZ ), 1.11 ng mL −1 sulfadiazine ( SD ), 1.15 ng mL −1 sulfapyridine ( SP ) and 5.27 ng mL −1 sulfamethoxazole ( SMX ). Desirable recoveries when detecting different spiked swine urine and milk samples were achieved ranging from 92.6% to 104.3% and from 61.1% to 81.6%, respectively.Conclusion The proposed immunoassay with the newly developed RabMAb is capable of detection of four SAs ( STZ , SD , SP and SMX ) with proven satisfactory performance and is applicable for routine large‐scale analysis in practical uses.© 2012 Society of Chemical Industry