| Summary: | 碩士 === 靜宜大學 === 食品營養學系 === 88 === A sandwich enzyme-linked immunosorbent assay ( ELISA ) for the indicator protein lactate dehydrogenase ( LDH ) was developed to determine endpoint heating temperature ( EPT ) of porcine muscle. Ground pork was placed in 10 by 75㎜ tubes and heated to temperatures between 64 and 74℃ at 2℃ intervals . Electrophoresis of meat extracts indicated a decrease in the intensity of 35 KDa band between 68 and 72℃. This protein was identified as LDH on the basis of molecular weight determination by SDS-PAGE, enzyme assay, LDH specific stain on native PAGE, and immunoblotting. Polyclonal antibodies ( PAb ) were raised in rats against porcine muscle LDH and a sandwich ELISA was developed using PAb for capture and biotinylated PAb for detection. Porcine muscle LDH PAb cross-reacted with bovine muscle and rabbit muscle LDH. The LDH content as measured by sandwich ELISA decreased in fresh/cured pork tender loin and ham heated between 68 and 72℃. Cold storage ( 4℃, 1wk ) and frozen storage ( -20℃, 1 mo ) did not have marked influence on sandwich LDH ELISA in differentiating EPT of porcine muscles between 68 and 72℃.
Key words : lactate dehydrogenase, ELISA, endpoint temperature, porcine muscle.
|
|---|
