Purification of Transthyretin by High Performance Affinity Chromatography from Human Plasma

Abstract

The partial purification of human transthyretin (TTR) by high performance affinity chromatography with the help of other separation techniques is described in the present paper. A new affinity medium was prepared with a monosized (ca. 10 microns particle size) macroporous resin as the support and thyroxine (T4) as the ligand. The purification of TTR was carried out in a few simple steps involving serum precipitation, anion exchange, Thyroxine affinity chromatography and gel filtration. The overall yield was 29% and the refined TTR contained less than 2% impurities as analyzed by RP-HPLC. When TTR was administrated to the culture medium DMEM of liver tumor strain SMMC-7721, a true inhibition of cell growth (ca. 50%) was observed as an actual decrease in cell number over time.

A Method for Purifying Native Transthyretin from Human Plasma

“…TTR has also been produced in the eukaryotic Pichia Pastoris system [27]. Lin et al [28] created a method consisting of serum precipitation, anion exchange, thyroxine affinity chromatography and gel filtration. The latest methodologies for TTR purification include: 1) salting out, anion and cation exchange chromatographies, preparative electrophoresis, and size chromatography [29]; 2) ammonium sulfate fractionation followed by urea/Sephadex G-100 chromatography and a combination of two dye-affinity chromatographic steps on reactive yellow and cibacron blue coupled to agarose columns [30] and 3) extraction of TTR fibrils followed by sequential gel filtration after solubilization in a solution of guanidine hydrochloride and covalent chromatography [31].…”