Navi Mumbai, India
Navi Mumbai, India

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Gnanasekar R.,Radiopharmaceuticals Programme | Nagvekar U.H.,Radiopharmaceuticals Programme | Sivaprasad N.,Radiopharmaceuticals Programme
Journal of Liquid Chromatography and Related Technologies | Year: 2010

We describe a two-step radioimmunoassay procedure (RIA) for the measurement of free thyroxine (fT4) in human serum. A commercial antibody with the affinity of 5.5 × 109 L/M was used for this study. The anti-T4 antibody was immobilized on the inner walls of the polystyrene tubes by passive adsorption through normal rabbit IgG and anti-rabbit IgG as immunobridges. The method developed uses very small amounts of antibody with minimum dilution of the sample. The assay covers the range of 0 to 87.5pmol/L with sensitivity of 0.9pmol/L. The intra-assay precisions were 6.3% and 7% at 6.7 and 20.7pmol/L, respectively, for 15 replicates. The inter-assay precision at 6.4 and 24.4pmol/L were 13% and 11.6%, respectively. The normal range established by analyzing 54 healthy volunteers was 15.3-24.7pmol/L and that of 69ambulatory subjects was 14.8-23.3pmol/L. Progressive dilution of euthyroid and hypothyroid samples up to 50-fold yielded virtually constant values, whereas a registered decrease in free T4 was observed with hyperthyroid samples. Free T4 estimated for 75 samples, including samples from 16 pregnant female subjects, correlated well with the values obtained by a reputed commercial kit with the linear correlation coefficient of 0.84. Copyright © Taylor & Francis Group, LLC.

Rasmi R.R.,Mangalore University | Shenoy K.B.,Mangalore University | Kadwad V.B.,Radiopharmaceuticals Programme | Sarnaik J.,Radiopharmaceuticals Programme | Somashekarappa H.M.,Mangalore University
Journal of Radioanalytical and Nuclear Chemistry | Year: 2015

The present study was oriented to develop, optimize and validate immunoradiometric assay for the measurement of C-peptide in human serum by employing magnetizable cellulose particles prepared by a novel method of wet grinding. Six commercial monoclonal antibodies raised against the different epitopes of C-peptide were evaluated for their compatibility as sandwich partners by “cross matching”. The developed one step inclusive assay with best matched combination of sandwich complex, “magnetizable cellulose particles linked capture antibody–C-peptide–125I labelled detector antibody” had an analytical sensitivity of 0.16 ng/ml and standard range of 0.3–30 ng/ml covering required physiological and diagnostic range. © 2015, Akadémiai Kiadó, Budapest, Hungary.

Rasmi R.R.,Mangalore University | Bhasker Shenoy K.,Mangalore University | Sarnaik J.,Radiopharmaceuticals Programme | Kadwad V.B.,Radiopharmaceuticals Programme | And 2 more authors.
Journal of Radioanalytical and Nuclear Chemistry | Year: 2014

We describe a convenient and flexible solid phase radioimmunoassay for human insulin employing magnetizable cellulose particles. Anti-porcine insulin antibody was covalently linked to magnetizable cellulose particles to form a stable and economical solid phase immunosorbent system. The tracer was prepared by radioiodinating insulin with 125I using Chloramine-T oxidation method. The analytical sensitivity of assay observed was 5.5 μIU/mL. Intra-assay and inter-assay variations were found to be <12 % along with analytical recovery of 93–109 %. The developed assay can be used for the routine analysis of clinical samples. In addition, concentration of the solid phase magnetizable immunosorbent can be easily varied as per the specific requirement for research purposes. © 2014, Akadémiai Kiadó, Budapest, Hungary.

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