Monobiotinylated relaxins

Biotinylated porcine relaxins were synthesized and purified to homogeneity. Native porcine relaxin was reacted with biotinyl-epsilon-aminohexanoic acid-N-hydroxysuccinimide ester and the resulting mixture was separated by either ion exchange chromatography on CM-cellulose at pH 5.5 or reversed-phase high performance liquid chromatography. All four possible monosubstituted relaxin derivatives, with substitutions in N alpha A1, N epsilon A7, N epsilon A16, and N epsilon B8, were obtained. All derivatives were fully biologically active and comparative circular dichroism spectroscopy showed no significant differences in their secondary structure.