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CLC number: S896.8

On-line Access: 2015-02-02

Received: 2014-08-17

Revision Accepted: 2014-09-21

Crosschecked: 2015-01-08

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Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Li-rong Shen

http://orcid.org/0000-0002-3197-9245

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Journal of Zhejiang University SCIENCE B 2015 Vol.16 No.2 P.155-166

http://doi.org/10.1631/jzus.B1400223


Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody


Author(s):  Li-rong Shen, Yi-ran Wang, Liang Zhai, Wen-xiu Zhou, Liang-liang Tan, Mei-lu Li, Dan-dan Liu, Fa Xiao

Affiliation(s):  Zhejiang Key Laboratory for Agro-Food Processing, Fuli Institute of Food Science, Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   shenlirong@zju.edu.cn

Key Words:  Freshness, Royal jelly, Major royal jelly protein 1 (MRJP1), Enzyme-linked immunosorbent assay (ELISA), High specific antibody


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Li-rong Shen, Yi-ran Wang, Liang Zhai, Wen-xiu Zhou, Liang-liang Tan, Mei-lu Li, Dan-dan Liu, Fa Xiao. Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody[J]. Journal of Zhejiang University Science B, 2015, 16(2): 155-166.

@article{title="Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody",
author="Li-rong Shen, Yi-ran Wang, Liang Zhai, Wen-xiu Zhou, Liang-liang Tan, Mei-lu Li, Dan-dan Liu, Fa Xiao",
journal="Journal of Zhejiang University Science B",
volume="16",
number="2",
pages="155-166",
year="2015",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1400223"
}

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%T Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody
%A Li-rong Shen
%A Yi-ran Wang
%A Liang Zhai
%A Wen-xiu Zhou
%A Liang-liang Tan
%A Mei-lu Li
%A Dan-dan Liu
%A Fa Xiao
%J Journal of Zhejiang University SCIENCE B
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1400223

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T1 - Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody
A1 - Li-rong Shen
A1 - Yi-ran Wang
A1 - Liang Zhai
A1 - Wen-xiu Zhou
A1 - Liang-liang Tan
A1 - Mei-lu Li
A1 - Dan-dan Liu
A1 - Fa Xiao
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DOI - 10.1631/jzus.B1400223


Abstract: 
Major royal jelly protein 1 (MRJP1), designated apalbumin 1, has been regarded as a freshness marker of royal jelly (RJ). A MRJP1-specific peptide (IKEALPHVPIFD) identified by bioinformatics analysis of homologous members of the major royal protein family was synthesized and used to raise polyclonal anti-MRJP1 antibody (anti-SP-MRJP1 antibody). Western blot analysis showed that anti-SP-MRJP1 antibody only reacted with MRJP1 in RJ. In contrast, the previously reported antibody against recombinant MRJP1 (anti-R-MRJP1 antibody) reacted with other members of MRJP family in RJ. enzyme-linked immunosorbent assay (ELISA) using anti-SP-MRJP1 antibody demonstrated that MRJP1 content in RJ stored at 40 °C significantly degraded by 37.3%, 55.9%, 58.0%, 60.6%, 65.7%, 72.7%, and 73.1% at 7, 14, 21, 28, 35, 42, and 49 d, respectively, when compared with MRJP1 content in fresh RJ (0 d). Optical density analysis of MRJP bands from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles demonstrated that the degradation of MRJP1, MRJP2, MRJP3, and MRJP5 in RJ was strongly and positively correlated with the period of storage (P<0.0001). Our results indicated anti-SP-MRJP1 antibody was highly specific for MRJP1, and ELISA using the antibody is a sensitive and easy-to-use method to determine the freshness and authenticity of RJ.

用高度特异性王浆主蛋白1多克隆抗体ELISA法检测蜂王浆新鲜度

目的:为蜂王浆主蛋白1(MRJP1)的快速检测和鉴别提供科学依据,为蜂王浆的质量控制提供技术 支持。
创新点:首次比较了MRJP1特异性多克隆抗体与MRJP1重组表达蛋白多克隆抗体对王浆主蛋白家族的免疫反应差异,验证了蜂王浆中MRJP1蛋白降解与保温时间的相关性,建立了以MRJP1作为蜂王浆新鲜度生物标志物的快速检测方法。
方法:通过蜂王浆主蛋白家族蛋白的氨基酸序列同源性分析,筛选出MRJP1的特异性多肽区域,进行人工合成,免疫兔子后取血清制备成特异性多克隆抗体。用蛋白质印迹法(Westernblot)检测了MRJP1特异性多克隆抗体与MRJP1重组表达蛋白多克隆抗体对王浆主蛋白家族的免疫反应。以新鲜蜂王浆为对照品,用MRJP1特异性抗体酶联接免疫吸附剂测定(ELISA)法和变性电泳胶灰度扫描法分别测定保温(40°C)7~49天的蜂王浆中MRJP1含量的变化,并进行了相关性 分析。
结论:MRJP1的特异性抗体对MRJP1蛋白具有专一的免疫识别特性,可特异性地检测代表蜂王浆新鲜度的MRJP1含量变化,并鉴别蜂王浆的真伪。

关键词:新鲜度;蜂王浆;王浆主蛋白1;ELISA;高度特异性

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