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Sandiwch ELISA for Detecting Bovine Blood in Ground Beef and Animal Feed

Title: A Sandiwch ELISA for Detecting Bovine Blood in Ground Beef and Animal Feed.
Name(s): Ofori, Jack Appiah, author
Hsieh, Yun-Hwa Peggy, professor directing thesis
Sathe, Shridhar K., committee member
Dorsey, Jodee, committee member
Moore, Mary Ann, committee member
Department of Nutrition, Food, and Exercise Science, degree granting department
Florida State University, degree granting institution
Type of Resource: text
Genre: Text
Issuance: monographic
Date Issued: 2006
Publisher: Florida State University
Place of Publication: Tallahassee, Florida
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Bovine plasma proteins are used as high quality ingredients in feed for farm animals and also as a binder or colorant in ground beef. Religious observance, as well as recent fears of epidemic bovine spongiform encephalopathy, calls for suitable methods to detect bovine blood in processed food and animal feed for regulatory purposes. Analytical methods are currently not available for this purpose because the available methods are neither bovine specific, tissue specific nor based on the detection of a thermal-stable analyte in blood. This research sought to develop a sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of bovine blood in human food and animal feed. Two bovine-specific monoclonal antibodies (Mab 6G12 and Mab 3D6) that bind to the same antigenic thermal-stable protein (60kDa) in heat-treated bovine blood were selected to construct the sandwich ELISA. Mab 6G12 was used as the coating antibody and biotinylated Mab 3D6 was used as the detecting antibody. Soluble proteins were extracted with 10 mM phosphate buffered saline (PBS, pH 7.2). This sandwich ELISA was bovine specific, with no cross reactivity to other bovine tissues or other common food additive proteins such as soy protein, egg albumin, and gelatin. The assay, however, showed trace cross-reaction with non-fat dry milk. The cross-reactivity can be eliminated when antibody dilution is increased. The assay had a detection limit of 0.1% of spray-dried bovine plasma in spray-dried porcine plasma and 0.5% whole bovine blood powder in spray- dried porcine plasma, with a sensitivity of 100%, specificity of 100% and an overall accuracy of 100%. Low levels (1% v/w) of bovine blood in raw and cooked beef were successfully detected by the sandwich assay. The assay is suitable for detecting bovine blood in animal feed as well as cooked and fresh ground beef because it is bovine specific, tissue specific and based on a thermo-stable bio-marker.
Identifier: FSU_migr_etd-2307 (IID)
Submitted Note: A Thesis Submitted to the Department of Nutrition, Food and Exercise Sciences in Partial Requirements for the Degree of Master of Science.
Degree Awarded: Summer Semester, 2006.
Date of Defense: June 30, 2006.
Keywords: Blood Infectivity, Transmissible Spongiform Encephalopathies, Prions In Blood
Bibliography Note: Includes bibliographical references.
Advisory Committee: Yun-Hwa Peggy Hsieh, Professor Directing Thesis; Shridhar K. Sathe, Committee Member; Jodee Dorsey, Committee Member; Mary Ann Moore, Committee Member.
Subject(s): Food
Persistent Link to This Record:
Owner Institution: FSU

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Ofori, J. A. (2006). A Sandiwch ELISA for Detecting Bovine Blood in Ground Beef and Animal Feed. Retrieved from