Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (5): 874-877.doi: 10.3969/j.issn.1673-8225.2011.05.027

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Establishment of a sandwich ELISA for RANTES detection and its application in rat small bowel transplantation

Kang Zhen-hua1, Wang Chun-yan2, Wang Wei-zhong3, Du Jian-jun3, Zheng Jian-yong3, Wang Lei1   

  1. 1Department of Colorectal and Anus Surgery, First Hospital, Jilin University, Changchun  130021, Jilin Province, China
    2Department of Immunology, Jilin Medical College, Jilin  132013, Jilin Province, China
    3Department of Gastrointestinal Surgery, Xijing Hospital, Fourth Military Medical University of Chinese PLA, Xi’an  710032, Shaanxi Province, China
  • Received:2010-07-01 Revised:2010-09-16 Online:2011-01-29 Published:2011-01-29
  • Contact: Wang Lei, Master’s supervisor, Professor, Department of Colorectal and Anus Surgery, First Hospital, Jilin University, Changchun 130021, Jilin Province, China Wanglei11@163.com
  • About author:Kang Zhen-hua☆, Doctor, Attending physician, Department of Colorectal and Anus Surgery, First Hospital, Jilin University, Changchun 130021, Jilin Province, China k_z_h75@163.com Wang Chun-yan☆, Doctor, Lecturer, Department of Immunology, Jilin Medical College, Jilin 132013, Jilin Province, China c_cy66@163. com Kang Zhen-hua and Wang Chun-yan contributed equally to this paper.

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Abstract:

BACKGROUND: RANTES and its receptors mediated cellular immunity are the essential components of acute allograft rejection, the dynamic observation on expression of RANTES in serum after small bowel transplantation maybe act as a relatively sensitive index for diagnosing acute allograft rejection.
OBJECTIVE: To establish a Sandwich ELISA for RANTES, utilize it in rat small bowel transplantation model and to discuss the possibility of early diagnosis by using ELISA in acute rejection.
METHODS: RANTES mAb was labeled by horseradish peroxidase and its discriminating epitopes was measured by competed ELISA. The sandwich ELISA kit was established sensitivity was detected. After that, the kit was used to measure RANTES in rat small bowel transplantation models. 
RESULTS AND CONCLUSION: In antibody-sandwich ELISA, the optimal capture mAb was No2 (5 mg/L), and the optimal anzyme-conjugate mAb was No4 (1: 800). The minimal RANTES antigen which could be detected by this method was 0.5 μg/L. The level of RANTES in serum was heightened obviously in heterogen rat small bowel transplantation group. A sensitive and specific antibody-sandwich ELISA was established for RANTES detection, which offers a new method to early diagnose acute rejection in small bowel transplantation.

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