Premium
The Purification of Human Urinary Kallikrein with Ion‐Exchange Radial Flow Membrane Chromatography
Author(s) -
Wang Hailin,
Li Tong,
Zou Hanfa,
Zhang Yukui,
Chao Julie,
Chao Lee
Publication year - 1996
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/(sici)1099-0801(199605)10:3<139::aid-bmc567>3.0.co;2-v
Subject(s) - chemistry , chromatography , kallikrein , ion exchange , membrane , ion , biochemistry , enzyme , organic chemistry
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.