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 ...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（antiSP-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.展开更多
基金supported by the Public Beneficial Scientific&Technical Plan of Zhejiang(No.2011C22039)the Important Scientific & Technical Plan of Zhejiang(No.2011C12023)+2 种基金the Important Scientific & Technical Innovation Project of Hangzhou(No.20131812A25)the Foundation of Fuli Institute of Food Science of Zhejiang University(No.KY201404)the National Natural Science Foundation of China(No.31271848)
文摘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（antiSP-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.
