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Development of radioimmunoassay for estimation of C-peptide in human serum

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Abstract

C-Peptide radioimmunoassay (RIA) is an invaluable aid in diagnosis of diabetes, insulinoma and hypoglycaemia. An RIA for C-peptide was developed by using a polyclonal antibody for human C-peptide raised in rabbits and validated using human serum samples. Tyrosyated C-peptide was radioiodinated to a specific activity of around 283 µCi/µg by conventional chloramine-T method. The detection limit of the assay (90% binding) was 0.43 ng/mL and the ED50 value was 8.5 ng/mL. The assay with the standard range of 0.3–18 ng/mL and low non-specific binding provide superior assay features. Separation of the bound from free complex was carried out using 22% polyethylene glycol. Intra-assay and inter-assay variations were found to be < 11.5% along with analytical recovery of 90–104%. No cross reactivity was observed with proinsulin, glucagon, somatostatin, insulin and pancreatic polypeptide.The developed assay can be used for the routine analysis of clinical samples.

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Acknowledgements

The authors are grateful to Board of Research in Nuclear Sciences (BRNS) for funding this work. The authors are thankful to Dr. N. Sivaprasad, Former Senior General Manager, BRIT for his support.

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Authors and Affiliations

  1. Department of Applied Zoology, Mangalore University, Mangalagangothri, Karnataka, 574199, India

    Rajan Radha Rasmi & K. Bhasker Shenoy

  2. Radiopharmaceuticals Programme, Board of Radiation and Isotope Technology (BRIT), Vashi Complex, Sector 20, Vashi, Navi Mumbai, 400703, India

    V. B. Kadwad & Jayula Sarnaik

  3. Centre for Application of Radioisotopes and Radiation Technology (CARRT), Mangalore University, Mangalagangothri, 574199, India

    Rajan Radha Rasmi & H. M. Somashekarappa

  4. Department of Biotechnology, PSG College of Arts and Science, Coimbatore, 641014, India

    Rajan Radha Rasmi

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  1. Rajan Radha Rasmi
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  2. V. B. Kadwad
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  3. Jayula Sarnaik
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  4. K. Bhasker Shenoy
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  5. H. M. Somashekarappa
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Correspondence to K. Bhasker Shenoy.

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Rasmi, R.R., Kadwad, V.B., Sarnaik, J. et al. Development of radioimmunoassay for estimation of C-peptide in human serum. J Radioanal Nucl Chem 327, 923–928 (2021). https://doi.org/10.1007/s10967-020-07536-4

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  • DOI: https://doi.org/10.1007/s10967-020-07536-4

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