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Monoclonal antibody-based enzyme immunoassay for detection of porcine plasma in fish surimi
Authors:Raja Mohd Hafidz Raja Nhari  Nur Fadhilah Khairil Mokhtar  Irwan Hanish  Muhajir Hamid  Muhammad Asraf Alif Mohamed Rashidi  Nur Maisarah Shahidan
Affiliation:1. Laboratory of Halal Science Research, Halal Products Research Institute, Universiti Putra Malaysia, Serdang, Malaysia;2. Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Malaysia
Abstract:Detection of porcine plasma using indirect ELISA was developed using mAb B4E1 for the prevention of their usage in human food that creates religious and health conflicts. The immunoassay has a CV < 20% and did not cross-react to other meat and non-meat proteins. The sensitivity of the assay is 0.25% (w/w) of porcine plasma in spiked raw and cooked fish surimi. The assay did not produce a false positive result for any of the commercial fish surimi tested that were not contain porcine plasma. Determination of a 60-kDa antigenic protein of porcine blood using Western blot confirmed its presence in the plasma fraction of the porcine blood. Further proteomic analysis involving liquid chromatography–tandem mass spectrometry (LC-MS/MS) revealed the 60-kDa protein to be porcine serum albumin.
Keywords:Porcine plasma  ELISA  monoclonal antibody  antigenic protein  fish surimi
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