title: High-specificity quantification method for almond-by-products, based on differential proteomic analysis.
abstract: A highly specific competitive enzyme-linked immunosorbent assay (ELISA) protocol has been developed to identify and classify almond products based on differential proteomic analysis. We applied two-dimensional electrophoresis to compare the differences between almond and apricot kernels to search for almond-specific proteins. The amino acid of apricot Pru-1 was sequenced and aligned to almond Pru-1. One peptide, RQGRQQGRQQQEEGR, which exists in almond but not in apricot, was used as hapten to prepare monoclonal antibody against almond Pru-1. An optimized ELISA method was established using this antibody. The assay did not exhibit cross-reactivity with the tested apricot kernels and other edible plant seeds. The limit of detection (LOD) was 2.5-100μg/g based on different food samples. The recoveries of fortified samples at levels of twofold and eightfold LOD ranged from 82% to 96%. The coefficients of variation were less than 13.0%. Using 7M urea as extracting solution, the heat-treated protein loss ratios were 2%, 5% and 15% under pasteurization (65°C for 30min), baking (150°C for 30min) and autoclaved sterilization (120°C for 15min), respectively.
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China.
authors.forename: Shiwei
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China.
authors.forename: Shifeng
authors.initials: S
authors.lastname: Wang
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China.
authors.forename: Jingmin
authors.initials: J
authors.lastname: Huang
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China.
authors.forename: Xintian
authors.initials: X
authors.lastname: Lai
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China.
authors.forename: Yegang
authors.initials: Y
authors.lastname: Du
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Liu
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China.
authors.forename: Bifang
authors.initials: B
authors.lastname: Li
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China.
authors.forename: Ronghu
authors.initials: R
authors.lastname: Feng
authors.affiliation: Shenzhen Academy of Metrology and Quality Inspection, Shenzhen 518102, PR China. Electronic address: Yangguowu126@126.com.
authors.forename: Guowu
authors.initials: G
authors.lastname: Yang
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D000068181:Prunus dulcis / Q000737:chemistry*
keywords: Almond; Enzyme-linked immunosorbent assay; Proteome; Quantitative analysis