Cell affinity chromatography

Yan and his colleagues used a stable isotope labeled proteome internal standard to analyse protein expression alterations during oxidative stress in breast epithelial cells. Affinity chromatography was combined with MS/MS in the proteomic analysis of human 06-methylguanine-DNA methyltransferase. These studies demonstrated that proteomics can elucidate protein expre-ssions associated with the pharmacological and toxic effects of an anticancer drug when integrated with other biochemical methods. 

In spite of the widespread utilization of affinity chromatography in cell separation, there are still a considerable number of problems to be solved. The most serious problem is that there is always a substantial fraction of cells that are non-specifically adsorbed on the matrix surface. The research on cell affinity chromatography done in the last decade seems to be more biased towards the improvement in operating conditions than to the development of specially designed matrices for cell separation as well as the characterization of immobilized proteins. Nevertheless, there is no doubt that further advances in affinity chromatography as an effective tool for cell separation virtually depend on the detailed understanding of the features of matrix materials and immobilized proteins, as well as their interacions at the interface. In this respect, cell affinity selection based on the specific interaction of cells with immobilized proteins on a solid-phase matrix is now a major area of interest in the field of biomaterials science. 

This paper briefly reviews the progress and present status of cell affinity selection with special emphasis on immuno-affinity chromatography, and then deals with our own studies on the development of new matrix materials specially designed for cell affinity chromatography. 

Development of tert-Amine Derivatized Matrices for Cell Affinity Chromatography... 

Summarizing the above description, the surface of polyamine graft copolymers with a definite amount of polyamine branches showed an extremely small quantity of non-specifically adsorbed lymphocytes. This advantageous characteristic of polyamine graft copolymers led us to utilize them as solid-phase matrices for cell affinity chromatography. 

Affinity chromatography of surface glycoproteins and immunoglobulin E receptors from rat basophilic leukaemia cells Affinity chromatography of membrane immunoglobulin M and murine lymphocyte immunoglobulin study of their affinities for the lectin... 

The most conventionally used macroscale affinity-based cell separation system is cell affinity chromatography (CAC). These systems can provide relatively high purity (>95 %) and high throughput (10-10 cells/h). CAC procedures can be divided into three consecutive steps loading,... 

Affinity chromatography using factor XII as ligand leads to purification of u-PAR rather selectively, with only trace quantities of cytokeratin 1 or gClqR present [K. Joseph and A. Kaplan, unpubl. observations]. It is of interest that none of these three proteins possesses a transmembrane domain but u-PAR has a phos-phatidylinositol linkage within the cell membrane. Nevertheless, each of them has been isolated from purified cell membranes and they have been demonstrated to exist within the cell membrane by immunoelectron microscopy [41] presumably... 

Protein affinity chromatography can be used for the separation of an individual compound, or a group of structurally similar compounds from crude-reaction mixtures, fermentation broths, or cell lysates by exploiting very specific and well-defined molecular interactions... 

The clearing effect of heparin on blood is associated with release of the triglyceride-hydrolyzing enzyme lipoproteinlipase (LPL) from the surface of endothelial cells.458,459 In addition to a number of apparently equivalent lipases from different tissues,459 heparin also releases a hepatic lipase.460,461 As suggested by the results of affinity-chromatography studies, this release is probably associated with binding of the polysaccharide to the enzyme. 62,463 Because other polyanions,464 including the... 

Purification of eIF4Efrom cell lysates for IEF eIF4E and associated proteins are isolated from cell extract by affinity chromatography m7GTP-Sepharose (as described later) and the beads were washed twice with 1 ml of lysis buffer. 18 p of m7GTP (100 pM) is added to the beads and incubated for 15 min at 4°. After centrifugation at 7000 rpm for 30 s, the supernatant is mixed with 7x sample buffer and urea (see previously), and loaded onto prefocused IEF gel. 

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