Development of immunochemical systems for detection of human skeletal troponin i isoforms

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Abstract

Troponin I (TnI), together with troponin T and troponin C (TnC), forms the troponin complex, a thin filament protein of the striated muscle that plays a key role in the regulation of muscle contraction. In humans, TnI is represented by three isoforms: cardiac, which is synthesised only in the myocardium, and fast and slow skeletal, which are synthesised in fast- and slow-twitch muscle fibres, respectively. Skeletal TnI isoforms can be used as biomarkers of skeletal muscle damage of various aetiologies, including mechanical trauma, myopathies, muscle atrophy (sarcopenia), and rhabdomyolysis. Unlike classical markers of muscle damage, such as creatine kinase or myoglobin, which are also present in other tissues, skeletal TnIs are specific for skeletal muscle. In this study, we developed a panel of monoclonal antibodies for immunochemical detection of skeletal TnI isoforms in Western blotting (sensitivity: 0.01-1 ng per lane), in immunohistochemistry, and in fluorescence immunoanalysis. Some of the designed fluorescence immunoanalyses enable the quantification of fast skeletal (limit of detection [LOD]=0.07 ng/mL) and slow skeletal TnI (LOD=0.1 ng/mL) or both isoforms (LOD=0.1 ng/ml). Others allow the differential detection of binary (with TnC) or ternary (with troponin T and TnC) complexes, revealing the composition of troponin forms in human blood.

About the authors

A. P. Bogomolova

Lomonosov Moscow State University; Hytest Ltd.

Author for correspondence.
Email: bogomolova.agnessa@yandex.ru

Faculty of Biology

Russian Federation, 119234 Moscow; Turku, Finland

I. A. Katrukha

Lomonosov Moscow State University; Hytest Ltd.

Email: bogomolova.agnessa@yandex.ru

Faculty of Biology

Russian Federation, 119234 Moscow; Turku, Finland

A. M. Emelin

Avtsyn Research Institute of Human Morphology of FSBSI “Petrovsky National Research Centre of Surgery”

Email: bogomolova.agnessa@yandex.ru
Russian Federation, 117418 Moscow

A. I. Zabolotsky

Lomonosov Moscow State University

Email: bogomolova.agnessa@yandex.ru

Faculty of Biology

Russian Federation, 119234 Moscow

A. V. Bereznikova

Lomonosov Moscow State University; Hytest Ltd.

Email: bogomolova.agnessa@yandex.ru

Faculty of Biology

Russian Federation, 119234 Moscow; Turku, Finland

O. S. Lebedeva

Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency

Email: bogomolova.agnessa@yandex.ru
Russian Federation, 119435 Moscow

R. V. Deev

Avtsyn Research Institute of Human Morphology of FSBSI “Petrovsky National Research Centre of Surgery”

Email: bogomolova.agnessa@yandex.ru
Russian Federation, 117418 Moscow

A. G. Katrukha

Lomonosov Moscow State University; Hytest Ltd.

Email: bogomolova.agnessa@yandex.ru

Faculty of Biology

Russian Federation, 119234 Moscow; Turku, Finland

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