title: Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody.
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.
authors: Zhejiang Key Laboratory for Agro-Food Processing, Fuli Institute of Food Science, Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310058, China.
authors: Li-rong
authors: LR
authors: Shen
authors: Yi-ran
authors: YR
authors: Wang
authors: Liang
authors: L
authors: Zhai
authors: Wen-xiu
authors: WX
authors: Zhou
authors: Liang-liang
authors: LL
authors: Tan
authors: Mei-lu
authors: ML
authors: Li
authors: Dan-dan
authors: DD
authors: Liu
authors: Fa
authors: F
authors: Xiao
mesh_terms: D000906:Antibodies / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005227:Fatty Acids / Q000032:analysis / Q000737:chemistry* / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology*; D019476:Insect Proteins / Q000276:immunology*; D008422:Materials Testing / Q000379:methods*; D000099219:Royal Jelly
chemical_list: D000906:Antibodies; D005227:Fatty Acids; D006023:Glycoproteins; D019476:Insect Proteins; D000099219:Royal Jelly; C509394:apalbumin 1, honeybee
keywords: Enzyme-linked immunosorbent assay (ELISA); Freshness; High specific antibody; Major royal jelly protein 1 (MRJP1); Royal jelly
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