Analysis of the major component of advanced oxidative protein products / 中华肾脏病杂志

ABSTRACT

Objective To design a method to characterize AOPPs. Methods Carbonyl groups were used as an oxidative index to link AOPPs and oxidized protein. Plasma-AOPPs were obtained by a series of preparation as follows. The native plasma was first determined for its protein contents followed by AOPPs level evaluation. Specimen was then washed with PBS and ultrafiltrated with an ultrafilter (10 000 cut-off membrane) to obtain clean plasma-AOPPs. A size-exclusion HPLC technique was used to verify which protein was oxidatively damaged. Fractions resulted from delipidation were also examined. Results The levels of AOPPs and total carbonyl groups in patient plasma were significantly higher than those in controls; both in native/delipidated plasma and CHC13-resulted precipitate. HPLC revealed that serum albumin presented highest carbonyl levels. It was an exclusive protein with statistically significant difference between controls and patients (patients vs. controls in nmol carbonyl/mg protein HSA 1.510?0.067 vs. 0.791?0.048, P