Wang H, Li T, Zou H, Zhang Y, Chao J, Chao L
National Chromatographic R. and A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Peoples Republic of China.
Biomed Chromatogr. 1996 May-Jun;10(3):139-43. doi: 10.1002/(SICI)1099-0801(199605)10:3<139::AID-BMC567>3.0.CO;2-V.
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.
