Development of a sensitive monoclonal antibody-based ELISA for the detection of sulfamethazine in cow milk, honey, and swine urine.
A specific monoclonal antibody (MAb) against sulfamethazine was produced with hybridoma technology. This assay shows very high sensitivity with IC50 of 0.4 ng/mL and LOD of 0.05 ng/mL when it was run in 0.02 mol/L PBS (pH 7.5). This MAb has shown high specificity to sulfamethazine, with little cross reactivity for sulfamerazine (5.27%) and sulfadimethoxypyrimidine (1.12%) and very low cross reactivity values for the other test compounds (≤0.1%). Sulfamethazine was spiked in milk, honey, and swine urine. After a simple extraction procedure the extracts at appropriate dilution were analyzed by ELISA. Satisfactory results were obtained by this assay, with average recoveries of 95-116% and coefficients of variation (CVs) of 5-9%. These results suggests that the MAb-based ELISA will be a feasible quantitative/screening method for sulfamethazine residue in real samples.
Medical Science College, Biotechnology Research Center, China Three Gorges University, Yichang, Hubei, China. y_t_b@hotmail.com
Tangbin
T
Yang
Xiangfu
X
Ren
Yuanbing
Y
Li
Faju
F
Chen
D000818:Animals; D000890:Anti-Infective Agents / Q000032:analysis / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D000918:Antibody Specificity; D002417:Cattle; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005240:Feasibility Studies; D005260:Female; D006722:Honey / Q000032:analysis*; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008892:Milk / Q000737:chemistry*; D008954:Models, Biological; D012680:Sensitivity and Specificity; D013418:Sulfamethazine / Q000032:analysis* / Q000302:isolation & purification; D013552:Swine / Q000652:urine*; D016482:Urinalysis / Q000379:methods
D000890:Anti-Infective Agents; D000911:Antibodies, Monoclonal; D013418:Sulfamethazine