THE PURIFICATION OF HUMAN URINARY KALLIKREIN WITH ION-EXCHANGE RADIALFLOW MEMBRANE CHROMATOGRAPHY

A method was developed for the purification of kallikrein from human u rine, The procedure consisted of three steps: ultradialysis, diethyl-( 2-hydroxypropyl) aminoethyl (QAE) ion exchange radial flow membrane ch romatography and affinity chromatography on aprotinin agarose, It is s imple and suitable for large-scale purification, The purified product was checked by SDS-PAGE and matrix-assisted laser desorption:ionizatio n mass spectrometery (MALDI), A single band with apparent molecular we ight (MW) 42,000 in SDS-PAGE and a single monomer peak with MW33,000 i n MALDI were observed, respectively, The biological activity tested by ELISA showed positive immunological identity of the purified product compared with human urinary kallikrein standard.