Establishment of double antibody sandwich ELISA for peanut allergenic protein Ara h 1

Authors

WANG Yao, College of Food and Bioengineering, Henan University of Science & Technology, Luoyang, Henan 471023 ,China
CHEN Xi, College of Food and Bioengineering, Henan University of Science & Technology, Luoyang, Henan 471023 ,China
WU Han-liang, Clinical Laboratory, No.989 Military Hospital, Joint Logistics Support Force of PLA, Luoyang,Henan 471031 , China
WEI Xing, Sanmenxia Inspection and Quarantine Center, Sanmenxia, Henan 472000 , China
ZHANG Guo-qing, Sanmenxia Inspection and Quarantine Center, Sanmenxia, Henan 472000 , China
ZHANG Shu-xia, Technology Center of Zhengzhou Customs, Zhengzhou, Henan 450002 , China
LIU Sheng-nan, Sanmenxia CustomsDistrict, Sanmenxia, Henan 472000 , China

Abstract

In order to establish a double antibody sandwich ELISA method for the rapid, sensitive and specific detection of peanut allergenic protein Ara h 1. Taking Ara h 1 murine monoclonal antibody (mAb) as capture antibody, and Ara h 1 rabbit polyclonal antibody (pAb) as detection antibody, the checkerboard method was used to optimize the antibody working concentration to establish the method, and the detection characteristics were identified, including sensitivity, specificity, accuracy and stability of the method. Results: The optimal working concentrations of mAb and pAb were 1∶10 000 and 1∶8 000 dilution, respectively. The linear range of the ELISA standard curve was 4~256 ng/mL, and the limit of detection was 4.16 ng/mL. The average recovery ranged from 85.9% to 94.5%, and there was no cross reaction with other common allergenic proteins. In addition, the detection results stable within 90 days under the condition of 4 ℃ in the dark and sealed. The established double antibody sandwich ELISA method is sensitive, specific, accurate and stable, which can be used for the rapid screening of peanut allergenic protein Ara h 1.

Publication Date

7-28-2020

First Page

59

Last Page

62,113

DOI

10.13652/j.issn.1003-5788.2020.07.012

Recommended Citation

Yao, WANG; Xi, CHEN; Han-liang, WU; Xing, WEI; Guo-qing, ZHANG; Shu-xia, ZHANG; and Sheng-nan, LIU (2020) "Establishment of double antibody sandwich ELISA for peanut allergenic protein Ara h 1," Food and Machinery: Vol. 36: Iss. 7, Article 12.
DOI: 10.13652/j.issn.1003-5788.2020.07.012
Available at: https://www.ifoodmm.cn/journal/vol36/iss7/12

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