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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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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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Reference

[1]Albert, S., Klaudiny, J., 2004. The MRJP/YELLOW protein family of Apis mellifera: identification of new members in the EST library. J. Insect Physiol., 50(1):51-59.

[2]Albert, S., Bhattacharya, D., Klaudiny, J., et al., 1999. The family of major royal jelly proteins and its evolution. J. Mol. Evol., 49(2):290-297.

[3]Antinelli, J.F., Zeggane, S., Davico, R., et al., 2003. Evaluation of (E)-10-hydroxydec-2-enoic acid as a freshness parameter for royal jelly. Food Chem., 80(1):85-89.

[4]AQSIQ (General Administation of Quality Supervision, Inspection and Quarantine of the People’s Republic of China), 2008. GB 9697-2008: The Quality Standard of Royal Jelly. National Standard of People’s Republic of China, China Standard Press (in Chinese).

[5]Bi?likova?, K., Simúth, J., 2010. New criterion for evaluation of honey: quantification of royal jelly protein apalbumin 1 in honey by ELISA. J. Agric. Food Chem., 58(15):8776-8781.

[6]Chen, C., Chen, S.Y., 1995. Changes in protein components and storage stability of royal jelly under various conditions. Food Chem., 54(2):195-200.

[7]Drapeau, M.D., Albert, S., Kucharski, R., et al., 2006. Evolution of the yellow/major royal jelly protein family and the emergence of social behavior in honey bees. Genome Res., 16(11):1385-1394.

[8]Fontana, R., Mendes, M.A., DeSouza, B.M., et al., 2004. Jelleines: a family of antimicrobial peptides from the royal jelly of honeybees (Apis mellifera). Peptides, 25(6):919-928.

[9]Fujii, A., Kobayashi, S., Kuboyama, N., et al., 1990. Augmentation of wound healing by royal jelly (RJ) in streptozotocin-diabetic rats. Jpn. J. Pharmacol., 53(3):331-337.

[10]Fujiwara, S., Imai, J., Fujiwara, M., et al., 1990. A potent antibacterial protein in royal jelly. J. Biol. Chem., 265(19):11333-11337.

[11]Furusawa, T., Rakwal, R., Nam, H.W., et al., 2008. Comprehensive royal jelly (RJ) proteomics using one- and two-dimensional proteomics platforms reveals novel RJ proteins and potential phospho/glycoproteins. J. Proteome Res., 7(8):3194-3229.

[12]Hanes, J., Simúth, J., 1992. Identification and partial characterization of major royal jelly protein of honey bee (Apis mellifera L.). J. Apic. Res., 31(1):22-26.

[13]Honda, Y., Fujita, Y., Maruyama, H., et al., 2011. Lifespan-extending effects of royal jelly and its related substances on the nematode Caenorhabditis elegans. PLoS ONE, 6(8):e23527.

[14]Howe, S.R., Dimick, P.S., Benton, A.W., 1985. Composition of freshly harvested and commercial royal jelly. J. Agric. Res., 24(1):52-61.

[15]Huesca, M., Sun, Q., Peralta, R., et al., 2000. Synthetic peptide immunogens elicit polyclonal and monoclonal antibodies specific for linear epitopes in the D motifs of Staphylococcus aureus fibronectin-binding protein, which are composed of amino acids that are essential for fibronectin binding. Infect. Immun., 68(3):1156-1163.

[16]Inoue, S., Koya-Miyata, S., Ushio, S., et al., 2003. Royal jelly prolongs the life span of C3H/HeJ mice: correlation with reduced DNA damage. Exp. Gerontol., 38(9):965-969.

[17]Kajiura, S., Yashiki, T., Funaoka, H., et al., 2008. Establishment and characterization of monoclonal and polyclonal antibodies against human intestinal fatty acid-binding protein (I-FABP) using synthetic regional peptides and recombinant I-FABP. J. Immunoassay Immunochem., 29(1):19-41.

[18]Kamakura, M., 2011. Royalactin induces queen differentiation in honeybees. Nature, 473(7348):478-483.

[19]Kamakura, M., Mitani, N., Fukuda, T., et al., 2001a. Antifatigue effect of fresh royal jelly in mice. J. Nutr. Sci. Vitaminol., 47(6):394-401.

[20]Kamakura, M., Suenobu, N., Fukushima, M., 2001b. Fifty-seven-kDa protein in royal jelly enhances proliferation of primary cultured rat hepatocytes and increases albumin production in the absence of serum. Biochem. Biophys. Res. Commun., 282(4):865-874.

[21]Kamakura, M., Fukuda, T., Fukushima, M., et al., 2001c. Storage-dependant degradation of 57-kDa protein in royal jelly: a possible marker for freshness. Biosci. Biotechnol. Biochem., 65(2):277-284.

[22]Klaudiny, J., Hanes, J., Kulifajova, J., et al., 2010. Molecular cloning of two cDNA from the heads of the nurse honey bee (Apis mellifera L.) for coding related proteins of royal jelly. J. Apicul. Res., 33(2):105-111.

[23]Kohno, K., Okamotom, I., Sanom, O., et al., 2004. Royal jelly inhibits the production of proinflammatory cytokines by activated macrophages. Biosci. Biotechnol. Biochem., 68(1):138-145.

[24]Lequin, R.M., 2005. Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA). Clin. Chem., 51(12):2415-2418.

[25]Li, J.K., Feng, M., Zhang, L., et al., 2008. Proteomics analysis of major royal jelly protein changes under different storage conditions. J. Proteome Res., 7(8):3339-3353.

[26]Li, Y., Chen, Y.X., Shen, L.R., et al., 2008. Prokaryotic expression of mrjp7 of Apis cerana cerana and preparation of its polyclonal antibody. J. Shanghai Jiaotong Univ. (Agric. Sect.), 26(2):109-113 (in Chinese).

[27]Majtan, J., Kovacova, E., Blikova, K., et al., 2006. The immunostimulatory effect of the recombinant apalbumin 1-major honeybee royal jelly protein-on TNFα release. Intern. Immunol. Pharmacol., 6(2):269-278.

[28]Malecová, B., Ramserb, J., O’Brienb, J.K., et al., 2003. Honeybee (Apis mellifera L.) mrjp gene family: computational analysis of putative promoters and genomic structure of mrjp1, the gene coding for the most abundant protein of larval food. Gene, 303:165-175.

[29]Matsui, T., Yukiyoshi, A., Doi, S., et al., 2002. Gastrointestinal enzyme production of bioactive peptides from royal jelly protein and their antihypertensive ability in SHR. J. Nutr. Biochem., 13(2):80-86.

[30]Messia, M.C., Caboni, M.F., Marconi, E., 2005. Storage stability assessment of freeze dried royal jelly by furosine determination. J. Agric. Food Chem., 53(11):4440-4443.

[31]Peixoto, L.G., Calabria, L.K., Garcia, L., et al., 2009. Identification of major royal jelly proteins in the brain of the honeybee Apis mellifera. J. Insect Physiol., 55(8):671-677.

[32]Sabatini, A.G., Marcazzan, G.L., Caboni, M.F., et al., 2009. Quality and standardisation of royal jelly. J. ApiProd. ApiMed. Sci., 1(1):1-6.

[33]Salazar-Olivo, L.A., Paz-Gonzales, V., 2005. Screening of biological activities present in honeybee (Apis mellifera) royal jelly. Toxicol. in Vitro, 19(5):645-651.

[34]Sambrook, J., Fritsch, E.F., Maniatis, T., 1989. Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.

[35]Schmitzová, J., Klaudiny, J., Albert, S., et al., 1998. A family of major royal jelly proteins of the honeybee Apis mellifera L. Cell. Mol. Life Sci., 54(9):1020-1030.

[36]Shen, L.R., Ding, M.H., Zhang, L.W., et al., 2010a. Expression of Acc-royalisin gene from royal jelly of Chinese honeybee in Escherichia coli and its antibacterial activity. J. Agric. Food Chem., 58(4):2266-2273.

[37]Shen, L.R., Zhang, W.G., Jin, F., et al., 2010b. Expression of recombinant AccMRJP1 protein from royal jelly of Chinese Honeybee in Pichia pastoris and its proliferation activity in an insect cell line. J. Agric. Food Chem., 58(16):9010-9017.

[38]Shinnick, T.M., Sutcliffe, J.G., Green, N., et al., 1983. Synthetic peptide immunogens as vaccines. Annu. Rev. Microbiol., 37(1):425-446.

[39]Simúth, J., 2001. Some properties of the main protein of honeybee (Apis mellifera) royal jelly. Apidologie, 32(1):69-80.

[40]Tamura, T., Fujii, A., Kuboyama, N., 1987. Antitumor effect of royal jelly. Folia Pharmacol. Japon., 89(2):73-80 (in Japanese).

[41]Tokunaga, K., Yoshida, C., Suzuki, K., et al., 2004. Antihypertensive effect of peptides form royal jelly in spontaneously hypertensive rats. Biol. Pharm. Bull., 27(2):189-192.

[42]Wu, L.M., Zhou, J.H., Xue, X.F., et al., 2009. Fast determination of 26 amino acids and their content changes in royal jelly during storage using ultra-performance liquid chromatography. J. Food Compost. Anal., 22(3):242-249.

[43]Yamaguchi, K., He, S., Li, Z., et al., 2013. Quantification of major royal jelly protein 1 in fresh royal jelly by indirect enzyme-linked immunosorbent assay. Biosci. Biotechnol. Biochem., 77(6):1310-1312.

[44]Zheng, H.Q., Hu, F.L., Dietemann, V., 2011. Changes in composition of royal jelly harvested at different times: consequences for quality standards. Apidologie, 42(1):39-47.

[45]Zheng, H.Q., Wei, W.T., Wu, L.M., et al., 2012. Fast determination of royal jelly freshness by a chromogenic reaction. J. Food Sci., 77(6):S247-S252.

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