The Purification of Human Urinary Kallikrein with Ion-Exchange Radial Flow Membrane Chromatography

✍ Scribed by Hailin Wang; Tong Li; Hanfa Zou; Yukui Zhang; Julie Chao; Lee Chao

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 393 KB
• Volume: 10
• Category: Article
• ISSN: 0269-3879
• DOI: 10.1002/(sici)1099-0801(199605)10:3<139::aid-bmc567>3.0.co;2-v

✦ Synopsis

A method was developed for the purification of kallikrein from human urine. The procedure consisted of three steps: ultradialysis, diethyl-(Z-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 hIW33.000 in hIALDI were observed, respectively. The biological activity tested by ELISA showed positive immunological identity of the purified product compared with human urinary kallikrein standard.