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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (09): 68-74    DOI: Q511
    
Development and Application of Heterologous ciELISA Kit for Rapid Detection of 19-Nortestosterone Residues
JIANG Jin-qing1,2,ZHANG Hai-tang1,WANG Zi-liang1,WANG Jian-hua 2,FAN Guo-ying1
1.College of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China
2.College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China
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Abstract  

To screen anti-Nortestosterone monoclonal antibody (NT mAb) and develop heterologous ciELISA kit. Balb/C mice were immunized with artificial antigen of NT-17-BSA and a strain of hybridoma cell named NT1-B7 was produced, which can stably secret NT mAb. The isotype of mAb was IgG1 /k, ascites titer was 0.64×105, affinity constant (Ka) was 4.31×109 L/mol and IC50 value determined by ciELISA was 0.55 ng/mL. Of all the analogous competitors, 2.2% and 1.6% cross-reactivity (CR) were observed for trenbolone and estradiol, respectively, while no CR was obtained for other compounds. With coating antigen of NT-3-OVA, heterologous ciELISA kit was developed, which limits of detection in pork, swine urine, beef and cattle urine were 0.24, 0.23, 0.22 and 0.17 ng/mL, respectively. The recoveries of NT spiked in the four matrix were in the range of 85%-110%, while intra- and inter-relative standard deviation (RSD) were in the range of 4.4%-21.8%. when stored at 4 ℃, the NT-Kit could preserve 6 months or more. The pH value of validation detection in matrix ranged from 6.5 to 7.5, and the program could save 30-90 min after the incubation process was optimized. With high sensitivity and specificity, this ciELISA kit could be advantageously utilized for the primary screening of sample matrix for presence of 19-NT residue.



Key words19-nortestosterone      Monoclonal antibodyci      ELISAHeterology      Rapid test kit     
Received: 10 May 2010      Published: 25 August 2010
Corresponding Authors: zi-liang wang     E-mail: wangzl_2008@yahoo.com.cn
Cite this article:

JIANG Jin-Qiang, ZHANG Hai-Tang, WANG Zi-Liang, WANG Jian-Hua, FAN Guo-Yang. Development and Application of Heterologous ciELISA Kit for Rapid Detection of 19-Nortestosterone Residues. China Biotechnology, 2010, 30(09): 68-74.

URL:

https://manu60.magtech.com.cn/biotech/Q511     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I09/68

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