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ZJUS-B
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology)  2015, Vol. 16 Issue (2): 155-166    DOI: 10.1631/jzus.B1400223
Articles     
Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody
Li-rong Shen, Yi-ran Wang, Liang Zhai, Wen-xiu Zhou, Liang-liang Tan, Mei-lu Li, Dan-dan Liu, Fa Xiao
Zhejiang Key Laboratory for Agro-Food Processing, Fuli Institute of Food Science, Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310058, China
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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.

Key words： Freshness      Royal jelly      Major royal jelly protein 1 (MRJP1)      Enzyme-linked immunosorbent assay (ELISA)      High specific antibody     
Received: 17 August 2014      Published: 02 February 2015
CLC:  S896.8  
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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
Cite this article:

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. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2015, 16(2): 155-166.

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http://www.zjujournals.com/xueshu/zjus-b/10.1631/jzus.B1400223     OR     http://www.zjujournals.com/xueshu/zjus-b/Y2015/V16/I2/155

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