Isolation and characterization of modified species of a mutated (Cys125 -Ala) recombinant human interleukin-2.

During purification of recombinant and mutated interleukin-2 (rhIL-2A125) by reversed-phase-high-performance liquid chromatography, more and less hydrophobic fractions named MHF and LHF, respectively are discarded due to the presence of some unidentified forms of rhIL-2Ala125. Using slow and linear gradients of acetonitrile, these fractions were further purified by RP-HPLC, analyzed by automatic Edman degradation, digested with trypsin and analyzed by electrospray ionization mass spectrometry. In all fractions, partial processing of the N-terminal Met residue was observed.