Premium
High performance ion exchange chromatography of human plasma lecithin: Cholesterol acyltransferase
Author(s) -
Holmquist Leif
Publication year - 1989
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/bmc.1130030612
Subject(s) - chemistry , chromatography , lecithin , elution , size exclusion chromatography , ion exchange , sterol o acyltransferase , acyltransferase , ion chromatography , cholesterol , lecithin—cholesterol acyltransferase , enzyme , biochemistry , apolipoprotein b , ion , lipoprotein , organic chemistry
Highly purified monomeric human plasma lecithin:cholesterol acyltransferase (LCAT), completely free of apolipoprotein D, has been chromatographed on a MonoQ HR 5/5 anion exchanger. LCAT eluted as symmetrical peaks after 12.8 min and 14.8 min at pH 5.0 and pH 6.0, respectively, using a linear NaCl gradient. The corresponding concentrations of NaCI effecting desorption of LCAT from the anion exchanger were 125 mM and 175 mM. At both pH values human serum albumin eluted earlier and was well separated from the enzyme. Rechromatography of LCAT in the eluates from these experiments at acid pH, on high performance gel filtration, demonstrated absence of aggregation. The nonspecific adsorption during anion exchange chromatography at pH 5.0 and pH 6.0 was negligible, as demonstrated by a linear relationship between injected amounts of LCAT and recorded peak areas for a 2–20 μg protein range. Zone immunoelectrophoresis assay indicated unaltered immunoreactivity of the eluted LCAT.