Apple F S, Ricchiuti V, Voss E M, Anderson P A, Ney A, Odland M
Department of Laboratory Medicine and Pathology, Hennepin County Medical Center and the University of Minnesota, Minneapolis 55415, USA.
Eur Heart J. 1998 Nov;19 Suppl N:N30-3.
The purpose of this study was to determine whether the two monoclonal anti-cardiac troponin T (cTnT) antibodies used in the second generation cTnT assay (capture Ab, M11.7; detection Ab, M7) would detect expression of cTnT isoforms in skeletal muscle from chronic renal disease patients. Skeletal muscle biopsies obtained from 45 chronic renal disease patients (as well as human heart muscles and normal human skeletal muscles) were prepared for Western blot analysis and blotted with the following anti-cTnT antibodies: M 11.7; M7; JS-2, Lakeland Biomedical; 13-11, Duke University) and anti-cTnI antibody JS-1. Using the M11.7 Ab, 20 of 45 renal skeletal muscles demonstrated one to three cTnT isoforms, MW34 39 kDa. These findings were confirmed by both the Lakeland and Duke antibodies. However the BM M7 antibody detected, in two of 45 muscles, only a protein with a MW of approximately 39 kDa. All four antibodies demonstrated equivalence in detection the 39 kDa cTnT isoform expressed in heart muscle. None detected any isoforms in normal skeletal muscle. A single cTnI isoform, MW 25 kDa, was detected by JS-1 only in normal adult myocardium. Based on the antibody configuration of the second generation cTnT assay, we conclude that while cTnT isoforms are expressed in human skeletal muscle obtained from chronic renal disease patients, if released into the circulation, they would not be detected.
本研究的目的是确定第二代心肌肌钙蛋白T(cTnT)检测中使用的两种单克隆抗心肌肌钙蛋白T抗体(捕获抗体,M11.7;检测抗体,M7)是否能检测慢性肾病患者骨骼肌中cTnT亚型的表达。从45例慢性肾病患者(以及人类心肌和正常人类骨骼肌)获取的骨骼肌活检样本用于蛋白质印迹分析,并用以下抗cTnT抗体进行印迹:M11.7;M7;JS - 2(莱克兰生物医学公司);13 - 11(杜克大学)以及抗心肌肌钙蛋白I抗体JS - 1。使用M11.7抗体时,45例肾性骨骼肌中有20例显示出一至三种cTnT亚型,分子量为34 - 39 kDa。莱克兰和杜克抗体均证实了这些发现。然而,BM M7抗体在45块肌肉中的两块中仅检测到一种分子量约为39 kDa的蛋白质。所有四种抗体在检测心肌中表达的39 kDa cTnT亚型时表现相当。在正常骨骼肌中均未检测到任何亚型。JS - 1仅在正常成人心肌中检测到一种分子量为25 kDa的单一心肌肌钙蛋白I亚型。基于第二代cTnT检测的抗体配置,我们得出结论,虽然cTnT亚型在慢性肾病患者的人类骨骼肌中表达,但如果释放到循环中,它们将无法被检测到。