Hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8) has been purified 23,000-fold from normal human erythrocytes. The purification includes affinity chromatography on a GMP column. The subunit molecular weight of the enzyme obtained from this purification is 24,000. The finding of four protein species after cross-linkage of the highly purified enzyme with dimethylsuberimidate, dimethyla...

The chemical synthesis of backbone deuterium labelled [r(CGCGAAU*U*CGCG)]2 (U* = [5'-2H]U) is described. An efficient purification procedure was developed using a polymeric reverse phase (PRP) HPLC column at 60 degrees C. This procedure provided pure RNA dodecamer in the multi-milligram quantities (39% overall yield) necessary for dynamics studies using solid-state deuterium NMR. The purificati...

Extraction and purification of Anthocyanin pigments from Black Raspberries (Rubus occidentalis) was successfully carried out. Homogeneous and fine powder was obtained from the liquid nitrogen blending process. After extraction and purification of the plant material, a colored solution was obtained. Depending on the nature of the predominant anthocyanin and its concentration, the coloration of t...

An efficient, scalable and high yielding process is developed for a neuromuscular blocking agent Rocuronium bromide 1 by studying various process parameters. The process involves simple work up and obviates column chromatography purification and usage of commercially unacceptable solvents.

Purpose: The refolding of recombinant human interferon α-2b is accompanied by low yield that could be due to high aggregation and multiplicity of steps in downstream processing. It is therefore essential to refold and purify the protein to simultaneously increase the production yield and reduce the required downstream steps. Methods: Inclusion bodies were dissolved in Tris-HCl buffer containing...

A T antigen preparation free of trypsin was obtained by application of CNBr-Sepharose linked to pure trypsin. Purification on an ion exchange chromatography column results in an electrophoretically homogeneous preparation of T protein.

P reparative chromatography is the dominant purification technique in the production of biological compounds, especially for bioseparations. The column used is central to the performance of the chromatographic step. To ensure robust and reproducible column performance, columns need to be qualified in an acceptable fashion before each use. Some of the critical elements of column qualification in...

BACKGROUND Alkyl hydroperoxide reductase (AhpC) of Helicobacter pylori is considered as a diagnostic antigen. Therefore, this antigen can be used to detect H. pylori infection by stool immunoassays such as ELISA. The aim of this study was to simplify the AhpC protein purification procedures. METHODS For whole cell protein extraction, the bacterial cells were ruptured by octly-β-D glucopyranos...

Choline acetyltransferase was demonstrated in nettles (Urtica dioica), peas (Pisum sativum), spinach (Spinacia oleracea), sunflower (Helianthus annuus) and blue--green algae by using a Sepharose--CoASH affinity column. The column effected a 1500-fold purification of the enzyme from nettle homogenates and was required for demonstrating activity in the other higher plants. Demonstration of the en...

In this paper, analysis of free nucleotides from mouseliver tissue during different day times has been described. Perchloric acid extract of mouse liver tissue was neutralized with tri-N-octylamine in trichlorotriflouroethane and after removal of ClO4(-), subjected to preliminary purification on a Cu(2+)-loaded column of Chelex 100. A high-pressure liquid chromatographic (HPLC) anion-exchange p...