|
|
|
| Screening of Hybridoma Lines of Monoclonal Antibody and Development of ciELISA Kit for Rapid Detection of Phenobarbital |
| WANG Zi-liang;ZHANG Gai-ping;ZHANG Hai-tang;YANG Yan-yan;WANG Yan-rong; CHAI Shu-jun |
1. College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China;
2. Henan Provincial Key Laboratory of Animal Immunology, Zhengzhou 450002, China |
|
|
|
|
Abstract Balb/c mice were immunized with BSA-pAPB and hybridoma lines secreted monoclonal antibody against phenobarbital (PB mAb) were generated with cell fusion and filtered by indirect ELISA and blocking ELISA. A ciELISA kit for detection PB (PB-Kit) was developed with PB mAb and its traits were tested. The results showed that three hybridoma lines were filtered and the best one was 3F6-C4, which secreted PB mAb with indirect ELISA titers of 1∶6.4×105 in ascites. PB mAb had a high affinity constant(Ka) with 1.96×1010 L/mol, a good sensitivity with an IC50 of 5.7 μg/L to PB, 12.4% cross-reactivity to barbiturate sodium and little or no cross-reactivity to other compounds. The PB-Kit had the linear detection range of 1.0 to 81 μg/L, the sensitivity of 0.75 μg/L and the detection limit of 1.0 μg/L. The recoveries of PB spiked in feed and swine urine were 85.8% and 91.3% respectively. The precision and accuracy of the assay determined by inter-assay and intra-assay coefficient variation was both below 15%. The PB-Kit possesses rapidity, sensitivity, specificity and briefness. That is proved to be used for the rapid detection of PB residues in animal food.
|
|
Received: 21 November 2005
|
|
|
|
Corresponding Authors:
ZHANG Gai-ping
|
|
|
|
|
|
|
