NDLI: A highly sensitive sandwich ELISA for the determination of glycomacropeptide to detect liquid whey in raw milk

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A highly sensitive sandwich ELISA for the determination of glycomacropeptide to detect liquid whey in raw milk

Content Provider	Springer Nature Link
Author	Chávez, Norma A. Jauregui, Juan Palomares, Laura A. Macías, Karla E. Jiménez, Mariela Salinas, Eva
Copyright Year	2012
Abstract	Milk processing industries and distributors have problems with adulteration of liquid milk by the addition of bovine cheese whey. Recently, the detection of fraudulent manipulation of milk with whey has focused on the identification of glycomacropeptide (GMP). Current non-immunological methods to detect GMP in dairy products are expensive and time-consuming or have low sensitivity. In this study, a novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of whey in raw milk was developed, using a polyclonal rabbit anti-GMP antibody. Calibration curves were constructed by analyzing raw milk standards containing different known concentrations of liquid cheese whey (0.02–20%). The method had a detection limit of 0.047% (v/v) and a quantification limit of 0.14% (v/v). The antibody showed high specificity and no cross-reaction with milk components (other than κ-casein) and was successful in detecting GMP in dairy commercial products. The recovery ratio was between 95.62% and 113.88% for all matrices tested. The intra-assay and interassay coefficients of variation were <6% and <7%, respectively. Finally, it can be stored for 3 months in the form of a ready-to-use kit, while maintaining its accuracy and reproducibility.摘要乳品生产企业和销售商们经常会遇到在液奶及乳制品中加入干酪乳清的问题。近年来,通过分析酪蛋白糖巨肽(GMP)的含量来确定乳中是否掺杂干酪乳清。非免疫法检测乳制品中GMP既昂贵又耗时,而且灵敏度低。目前,开发了一种新的夹心酶联免疫(ELISA)法实现了原料乳中乳清掺假问题的定性和定量检测。该方法以多克隆兔抗酪蛋白糖巨肽作为抗体,以已知浓度的液态干酪乳清(0.02–20%)为标准品建立校正曲线。方法的检测限为0.047% (v/v),定量限为0.14% (v/v)。抗体具有专一性,与乳中其它成分(κ-酪蛋白除外)没有交叉反应,能够成功的用于乳制品中酪蛋白糖巨肽的检测。对实验样品检测的检测回收率在95.62% ∼ 113.88%。批间和批内变异系数分别小于6%和7%。该抗体在三个月内,其准确度和重复性保持不变,并且可以随时取用。
Starting Page	121
Ending Page	132
Page Count	12
File Format	PDF
ISSN	19585586
Journal	Dairy Science & Technology
Volume Number	92
Issue Number	2
e-ISSN	19585594
Language	English
Publisher	Springer-Verlag
Publisher Date	2012-01-18
Publisher Place	Paris
Access Restriction	Subscribed
Subject Keyword	Glycomacropeptide Polyclonal antibodies Cheese whey Milk ELISA Agriculture Microbiology Food Science
Content Type	Text
Resource Type	Article
Subject	Biochemistry Food Science

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