Antibody development and enzyme-linked immunosorbent assay for the protein marker lactate dehydrogenase to determine safe cooking end-point temperatures of turkey rolls

Proteins as Potential Endpoint Temperature Indicators for Ground Beef Patties

“…Meat (25g) from the center of each patty was homogenized with 3 volumes (w/v) of 0.15M NaCl, 0.01M Na phosphate buffer, pH 7.2 in a Waring Blendor for 90 sec, and centrifuged at 16000 ϫ g for 30 min at 4ЊC (Wang et al, 1992). The supernatant was filtered through Whatman No.…”

“…Protein concentration of extracts was determined using the biuret method with BSA as standard (Gornall et al, 1949). Extractable protein was calculated as described by Wang et al (1992).…”

“…Lactate dehydrogenase and BSA were identified by Western blotting. Protein standards (LDH or BSA) and patty extract proteins were transferred electrophoretically (1 hr at 100V) from the SDS-PAGE gel to a nitrocellulose membrane (0.45 um, Schleicher and Schuell, Keene, NH) in a Mini Trans-Blot unit (Bio-Rad) using 25 mM Tris, 192 mM glycine, and 20% (v/v) methanol buffer (pH 8.3) as described by Wang et al (1992), except that 3.0% (w/v) gelatin in PBS was used as blocking agent for BSA.…”

“…Acid phosphatase (Kormendy et al, 1987(Kormendy et al, , 1992 and pyruvate kinase (Davis et al, 1988) were potential EPT indicators in canned hams and a model pork product, respectively. Lactate dehydrogenase (LDH) is a potential EPT indicator in bovine (Collins et al, 1991a;Stalder et al, 1991), porcine (Collins et al, 1991b) and poultry (Wang et al, 1992) muscle. Antibodies to poultry LDH were used to develop an immunoassay to verify EPT in poultry products (Wang et al, 1992;Abouzied et al, 1993).…”

“…Lactate dehydrogenase (LDH) is a potential EPT indicator in bovine (Collins et al, 1991a;Stalder et al, 1991), porcine (Collins et al, 1991b) and poultry (Wang et al, 1992) muscle. Antibodies to poultry LDH were used to develop an immunoassay to verify EPT in poultry products (Wang et al, 1992;Abouzied et al, 1993). Smith et al (1991) monitored changes in the endogenous enzyme activity of raw and processed (60, 70, 80ЊC) ground beef in a model system and reported that several enzymes might be used to verify EPT in beef prod- ucts.…”