Evaluation of a direct prorenin assay making use of a monoclonal antibody directed against residues 32-39 of the prosegment

J Hypertens. 2011 Nov;29(11):2138-46. doi: 10.1097/HJH.0b013e32834b1978.

Abstract

Background: Prorenin is an early marker of microvascular complications in diabetes. However, it can only be measured indirectly (following its conversion to renin), with a renin immunoradiometric assay (IRMA). Unfortunately, treatment with a renin inhibitor interferes with this assay, because renin inhibitors induce a conformational change in prorenin, thereby allowing its detection as renin.

Methods: We evaluated Molecular Innovation's new direct prorenin ELISA, which makes use of an antibody that recognizes an epitope near prorenin's putative cleavage site (R 43 L 44), thus no longer requiring prorenin activation. Plasma samples of 41 diabetic individuals treated with aliskiren (renin inhibitor) or irbesartan were tested. Semi-purified recombinant prorenin was used as standard, because the ELISA standard yielded approximately 10-fold lower values in the renin IRMA following its conversion to renin.

Results: The ELISA detected prorenin levels that were identical to those determined by the IRMA in untreated and irbesartan-treated individuals. Yet, it yielded higher prorenin levels in aliskiren-treated individuals. Aliskiren, at levels reached in plasma during treatment, did not interfere with the ELISA, but allowed the detection of up to 20-30% of prorenin as renin in the IRMA, thereby resulting in a significant overestimation of renin and an underestimation of prorenin. The ELISA rendered results within 2 h and did not require a pretreatment period of several days to convert prorenin to renin.

Conclusion: The new direct assay allows rapid prorenin detection, is not hampered by aliskiren when used at clinically relevant doses, and might be used to identify diabetic patients developing retinopathy and/or nephropathy.

MeSH terms

  • Aged
  • Antibodies, Monoclonal / chemistry*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Epitopes / chemistry
  • Female
  • Humans
  • Immunoradiometric Assay / methods
  • Kidney Diseases / pathology
  • Male
  • Microcirculation
  • Middle Aged
  • Protein Structure, Tertiary
  • Renin / blood*
  • Renin / chemistry
  • Time Factors

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • Renin