Recombinant Expression and Purification of Human Cystatin C Biomarker in Escherichia coli for Prediagnostic Renal Disorders

Introduction: chronic kidney disease is a worldwide health problem and glomerular filtration rate (gfr) is the most frequently used criteria in the assessment of rental function. cystatin c as a member of type 2 cystatin superfamily and cysteine-protease inhibitor is found in high concentrations in all biological fluids. this study is aimed to study cystatin c as a potent biomarker for clinical measurement of renal disorders and other diseases. materials and methods: in this study, the human cystatin c construct was analyzed by bioinformatics software. it was cloned and expressed to produce an appropriate antigen for anti-cystatin c (anti-cys c) obtained from mice balb/c as a crucial point in the improvement of an enzymelinked immunosorbent assay (elisa) method. serum samples were given from 32 hospitalized patients with renal failure and cardiovascular disease and non-hospitalized patients were tested by elisa method using anti-cys c obtained from mice balb/c. results: our findings indicated 0.36-2.4 mg/l as the best conclusion for antigen cystatin c in patients’ sera and 1/50 dilution for anti-cys c obtained from mice balb/c and showed a relationship between patients with high creatinine and high concentration of cystatin c. in case of five cardiovascular disease patients with normal upper limit of creatinine we obtained cystatin c lower than kidney failure and raising of cystatin c in 6 patients with increased tsh were seen. conclusions: polyclonal anti-cys c antibodies were obtained through the immunization of balb/c mice can be employed as an anti-cys c in elisa for diagnosis of some renal dysfunction.