Quantification of Relaxin-2 Connecting Peptide (Pro-RLX2) in Human Blood Samples

Abstract

Abstract Background The peptide hormone relaxin-2 is implicated in diverse physiological and pathophysiological processes. Several assays are available for quantification of human relaxin-2, but because stability of the mature peptide in serum is limited, measurement of the more stable connecting peptide (pro-RLX2) might be beneficial. Methods Pro-RLX2 was measured in a sandwich immunoluminometric assay using 2 monoclonal antibodies. The concentration of pro-RLX2 was detected in healthy pregnant (n = 100) and healthy male and nonpregnant female (n = 81) subjects and compared with the concentration of mature relaxin-2 in a subset of samples. Results The pro-RLX2 immunoassay has an analytical and functional assay sensitivity (FAS) of 1.59 pmol/L and 1.7 pmol/L, respectively. The analyte is stable in EDTA plasma samples for 8 days at room temperature, dilutes in a linear fashion, and recovery was 103%. The assay system is not biased by common interfering substances. Measurement of 80% of plasma samples from healthy males and females is below the FAS {median 1.49 pmol/L [interquartile range (IQR) of 0.925–2.14 pmol/L]}, and no concentration difference between male and nonpregnant female plasma samples was observed. The median plasma concentration in healthy pregnant women is increased up to 562 pmol/L (IQR 341–789 pmol/L). During pregnancy, pro-RLX2 concentrations decrease with increasing gestation. The correlation coefficient with the R&D assay for mature relaxin-2 was 0.96 (P < 0.0001). Conclusion Pro-RLX2 is stable in plasma of healthy individuals. Although samples of pregnant women are reliably measurable, most samples from healthy nonpregnant women and men are below the detection limit. Determination of pro-RLX2 concentrations might indicate rate of synthesis of relaxin-2 during pregnancy and therapeutic application of recombinant relaxin (Serelaxin).