The purification of human urinary kallikrein with ion-exchange radial flow membrane chromatography.

H Wang, T Li, H Zou, Y Zhang, J Chao, L Chao

Index: Biomed. Chromatogr. 10(3) , 139-43, (1996)

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Abstract

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