A skeletal muscle troponin T specific ELISA based on the use of an antibody against the soluble troponin T (16-31) fragment

被引:11
作者
Tsitsilonis, OE
Stoeva, S
Echner, H
Balafas, A
Margomenou, L
Katsoulas, HL
Troy, DJ
Voelter, W
Papamichail, M
Lymberi, P
机构
[1] Hellenic Pasteur Inst, Biochem Dept, Immunol Lab, Athens 11521, Greece
[2] St Savas Canc Hosp, Dept Immunol, Athens 11522, Greece
[3] Univ Tubingen, Inst Physiol Chem, Abt Phys Biochem, D-72076 Tubingen, Germany
[4] Natl Food Ctr, Dublin 15, Ireland
关键词
muscle troponin T; competitive ELISA; troponin T antibodies; troponin T peptides;
D O I
10.1016/S0022-1759(02)00186-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Proteolytic degradation of muscle, which occurs post-mortem as part of the meat-ageing process, results in the production of protein fragments. In beef, degradation of skeletal muscle troponin T (TnT) results in the generation of a 16-residue long peptide (TnT (16-31)), identified in trichloroacetic acid (TCA) soluble extracts. We report the development of a competitive enzyme-linked immunosorbent assay (ELISA) for the quantification of TnT (16-31), using polyclonal antibodies raised against synthetic TnT (16-31). The ELISA procedure is based on inhibition of binding of the antibodies to immobilised TnT (16-31) by TnT (16-31) present in solution. Its useful range is 30 pmol to 2 nmol TnT (16-31)/ml. Quantification of TnT (16-31) in TCA muscle extracts showed that its concentration was enhanced with ageing. Moreover, a correlation between TnT (16-31) levels and meat tenderness was observed. The ELISA developed herein may prove advantageous for future use at the research and industrial level. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:141 / 148
页数:8
相关论文
共 21 条
[1]  
CASSERLY U, 1998, P 44 ICOMST BARC SPA, V2, P726
[2]  
ETHERINGTON DJ, 1991, ENZYMES FOOD PROCESS, P128
[3]  
Kaiser T, 1997, Z NATURFORSCH B, V52, P281
[4]  
Kalichman S.C., 1990, ANN SEX RES, V3, P129, DOI DOI 10.1111/J.1745-4573.1990.TB00360.X
[5]  
Koohmaraie M, 1996, Meat Sci, V43S1, P193, DOI 10.1016/0309-1740(96)00065-4
[6]   EFFECTS OF POSTMORTEM STORAGE ON MUSCLE PROTEIN-DEGRADATION - ANALYSIS BY SDS-POLYACRYLAMIDE GEL-ELECTROPHORESIS [J].
KOOHMARAIE, M ;
KENNICK, WH ;
ELGASIM, EA ;
ANGLEMIER, AF .
JOURNAL OF FOOD SCIENCE, 1984, 49 (01) :292-293
[7]   A HIGH-INCIDENCE OF CROSS-REACTIVE IDIOTYPES AMONG MURINE NATURAL AUTOANTIBODIES [J].
LYMBERI, P ;
DIGHIERO, G ;
TERNYNCK, T ;
AVRAMEAS, S .
EUROPEAN JOURNAL OF IMMUNOLOGY, 1985, 15 (07) :702-707
[8]   Effects of freezing on the proteolysis of beef during storage at 4 degrees C [J].
Nakai, Y ;
Nishimura, T ;
Shimizu, M ;
Arai, S .
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1995, 59 (12) :2255-2258
[9]   The origin of the 30 kDa component appearing during post-mortem ageing of bovine muscle [J].
Negishi, H ;
Yamamoto, E ;
Kuwata, T .
MEAT SCIENCE, 1996, 42 (03) :289-303
[10]   RELATIONSHIP OF MYOFIBRIL FRAGMENTATION INDEX TO MEASURES OF BEEFSTEAK TENDERNESS [J].
OLSON, DG ;
PARRISH, FC .
JOURNAL OF FOOD SCIENCE, 1977, 42 (02) :506-509