Preservation of Biological LH and FSH Activity after Application on HPLC. Comparison between Cation-Exchange and Reverse-Phase High Performance Liquid Chromatography

Abstract

A cation-exchan e high-performance l i q u i d chromatography procedure i s described !or the separation o f b i o a c t i v e l u t r o p i n from f o l l i t r o p i n i n a human u r i n a r y gonadotropin standard p r e p a r a t i o n ( 1 s t IRP 70/45). The r e s u l t s c l e a r l y demonstrate t h a t recovered p r o t e i n r e t a f n e d most o f i t s b i o l o ical, a c t i v i t y f o l l o w i n g t h e chromatography. Due t o a l a c k o f e a f i c i e n t and quick methods f o r p u r i f y i n g p i t u i t a r o r u r i n a r y gonadotro ins. t h i s method could prove t o be valuabfe f o r t h i s purpose. TRis

Reconstitution of native-like nucleosome core particles from reversed-phase-HPLC-fractionated histones

“…In most of these, trifluoroacetic acid in the range 0.1-0.3 % (v\v) is used in the mobile phase and acetonitrile (in a continuous or stepwise gradient) is used as an organic modifier. The use of high concentrations of acetonitrile, which is known to be very destructive towards the secondary and tertiary structure of proteins [14,15], raises serious concerns and has restricted the use of RP-HPLC-purified histones for chromatin reconstitution experiments. In the present paper we address this concern and show that indeed RP-HPLC-purified histones can be used in the reconstitution of native-like nucleosome core particles regardless of the biological source or the method of extraction of the histones (acid or salt) before their HPLC purification.…”