Mechanisms
Recombinant cynomolgus monkey CETP is expressed and purified 176-fold from CHO cells for structural study (J Protein Chem 1995)
Original title: Expression and purification of recombinant cynomolgus monkey cholesteryl ester transfer protein from Chinese hamster ovary cells
The gene for cynomolgus monkey CETP was expressed in serum-free Chinese hamster ovary cell culture supplemented only with 2 micrograms/ml insulin, with cell growth aided by immobilizing the cells in alginate beads. A three-step affinity-chromatography protocol using Reactive Red 120 and concanavalin A Sepharose 4B purified recombinant CETP 176-fold with a 60% final yield, typically yielding 3.4 mg of protein from 1700 ml of media. SDS-PAGE showed a single broad band of 68,000 to 74,000 molecular weight that immunoreacted on Western blot, and amino acid analysis and protein sequencing matched the theoretical composition and sequence of cynomolgus CETP, establishing a source of purified recombinant CETP for further structural and functional study.
Original abstract
Cholesteryl ester transfer protein (CETP) mediates the transfer of cholesteryl ester from high- and low-density lipoproteins to triglyceride-rich lipoproteins, and reciprocally mediates triglyceride transfer. The gene for cynomolgus monkey CETP was expressed in serum-free CHO culture with 2 micrograms/ml insulin as its only exogenous protein supplement. Cell growth was facilitated by immobilizing the CHO cells in alginate beads. Recombinant CETP (rCETP) was purified 176-fold with a three-step protocol resulting in a 60% final yield as measured by a fluorescent CETP activity assay. Typically, 3.4 mg of rCETP was purified from 1700 ml of media by affinity-gel chromatography involving Reactive Red 120 (RR120) followed by concanavalin A Sepharose 4B and rechromatography on RR120. SDS-PAGE shows a single broad band of M(r) ranging from 68,000 to 74,000 which immunoreacts in Western blot analysis. Amino acid analysis and protein sequencing of the purified protein agree with the theoretical amino acid composition and sequence of cynomolgus CETP.
Summary written by cetpinhibition.org from the published abstract; figures as published. Page updated 18 August 2026. Methods.