CH2C12. The active fractions (CC14 and CH2C12, 300mg) were pooled and subjected to a flash silica column (CHCI3 : MeOHstep gradient). The active fraction (52 mg) eluted with 95:5 CHC13:MeOHand was subjected to a second flash silica column (hexanes : EtOAc step gradient). Final purification was obtained by normal phase HPLC (Sigma-Aldrich Nucleosil Silica 5 ,am, 25.0 cmX4.6 mm) with a solvent sy...

Column liquid chromatography (CLC) is a widely used unit operation in biotech and chemical process engineering for purification of target products (e.g. proteins for pharmaceutical agents) from unwanted substances. An aqueous solution containing the mixture of molecules is forced to flow through the chromatography column which is filled with porous particles (beads). While transport through the...

Recombinant plasmid DNA has been used to purify complementary cDNA by hybridization using a modification of sulfhydryl-Sepharose chromatography described by Dale and Ward ((1975) Biochemistry 14, 2458). Plasmid DNA containing cloned mouse globin or immunoglobulin sequences was mercurated and hybridized in solution to unpurified cDNA. The resulting hybrids were passed over a sulfhydryl-Sepharose...

its purification a benzoylated diethylaminoethyl cellulose column (7), after which we passed the preacylated tRNAsi^c: over an RPC-II column. This last column is described in Chart 1. The Chromatographie profile of the phenylalanine acceptor activity presented 3 distinct peaks. Those indicated as Phe 1 and Phe 2 were also normally present in tRNAs^a 0); Phe X is a new tRNA that we first observe...

A small, semi-preparative C(4) RP-HPLC column was used to set up the conclusive laboratory-scale purification of Chinese hamster ovary-derived human thyrotropin (hTSH), after a preliminary concentration-purification of an extremely dilute and poorly ( approximately 0.6 microg hTSH/mL; mass fraction=0.35%) conditioned medium on a cation exchanger. Several fractions of this eluate were repeatedly...

For clinical purposes, pure protein and identification of carbohydrate structure from recombinant erythropoietin are needed. Purification was done by Immobilized Metal Affinity Chromatography (IMAC) column charged with Ni (His-Trap affinity chromatography) and continued with gel filtration chromatography column to get purer protein. The carbohydrate group which is oligosaccharide from the resul...

Purification and PCR amplification procedures for DNA extracted from environmental samples (soil, compost, and river sediment) were improved by introducing three modifications: precipitation of DNA with 5% polyethylene glycol 8000 (PEG) and 0.6 M NaCl; filtration with a Sepharose 4B–polyvinylpolypyrrolidone (PVPP) spin column; and addition of skim milk (0.3% w/v) to the PCR reaction solution. H...

A simplified and efficient method is developed for the large-scale purification of the secoisolariciresinol diglucoside (SDG) from defatted flaxseed after aqueous ethanol extraction. Extractant from defatted flaxseed with aqueous ethanol is hydrolyzed with basic solution, concentrated under vacuum, subjected to Sephadex LH-20 column chromatography, and eluted with aqueous ethanol of different c...

Oligonucleotides are synthesized and purified in a single column filled with a mixture of nucleoside-loaded and underivatized, high-cross link, non-swelling polystryene beads. A new oligonucleotide production system has been developed to completely automate-the entire process of sequence entry, phosphoramidite synthesis, cleavage, deprotection, purification, quantitation, and sample collection....