A skeletal muscle troponin T specific ELISA based on the use of an antibody against the soluble troponin T (16-31) fragment.
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.
Tsitsilonis
Ourania E
OE
Department of Immunology, St Savas Cancer Hospital, 171 Alexandras Ave, 11522 Athens, Greece.
Stoeva
Stanka
S
Echner
Hartmut
H
Balafas
Apostolos
A
Margomenou
Lila
L
Katsoulas
Haralabos L
HL
Troy
Declan J
DJ
Voelter
Wolfgang
W
Papamichail
Michael
M
Lymberi
Peggy
P
D000818:Animals; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D018482:Muscle, Skeletal / Q000737:chemistry*; D010446:Peptide Fragments / Q000276:immunology*; D011817:Rabbits; D020107:Troponin T / Q000276:immunology*
D010446:Peptide Fragments; D020107:Troponin T