A novel diagnostic method for Duchenne Muscular Dystrophy using m-calpain as a quantitative marker: Biochemical marker for DMD diagnostics
Journal Title: Journal of Medical Science And clinical Research - Year 2014, Vol 2, Issue 1
Abstract
Calcium activated neutral protease-2 (m-Calpain) is a prominent catalytic protein found at the cellular level of human placenta. Earlier investigations stated its involvement in duchenne muscular dystrophy, which made us interested to isolate and purify m-Calpain with high yield to raise polyclonal antibodies. Here, we discussed indigenously developed reliable methodology by optimizing dependent parameters to isolate and purify m-Calpain with more yield. By currently developed methodology, significant yield of m-Calpain has been isolated and purified i.e 4-5 mg /1000g placenta tissue, which consists of 36.57% of total recovered protein. We have raised specific polyclonal antibodies against m-calpain in albino rabbits and serologically confirmed the purified enzyme by double diffusion and electrophoresis. Through development of this methodology, it enabled us to develop the cost-effective Quantitative ELISA (Q-ELISA) to identify Duchenne Muscular Dystrophy (DMD) patients and true carriers. Through Q-ELISA test we have found out that elevated levels of m-Calpain in platelets i.e 17.1±0.6 ng/ml in true carriers, 16.5±4.5 ng/ml in patients, 1.7±0.3 ng/ml in control males & 1.8±0.8 ng/ml in control females.
Authors and Affiliations
Dr. Mohana Rao V
Keywords
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