H Affinity Chromatography

Scouten, W.H., Affinity Chromatography Bioselective Adsorption on Inert Matrices, Wiley, New York, 1981. 

Schott, H., Affinity Chromatography, Chromatographic Science Series Vol. 27, Marcel Dekker, Inc., New York, 1984. 

Fig. 21 Chitin binding of 6xHis-tagged resilin with chitin-binding domain (6 H-resChBD) as compared to 6xHis-tagged resilin without chitin-binding domain (6 H-res). T total protein after affinity chromatography purification, B bound protein eluted from chitin beads, UB unbound protein. Reproduced from [187] with permission from The American Chemical Society, copyright 2009...

Candidates for the renal brush border Na /H exchanger transport protein identified by covalent labeling, affinity chromatography, or other methods... 

Huot et al. [38] used affinity chromatography to identify and partially purify an amiloride-binding protein with characteristics of the renal brush border Na /H exchanger. The high-affinity amiloride analog A35 (5-A-(3-aminophenyl)amiloride) was coupled to Sepharose CL-4B through a triglycine spacer. Rabbit renal brush border membranes were solubilized with 0.6% Triton X-100, incubated with the... 

In a preliminary report, Ross et al. [40] used affinity chromatography to identify a putative bovine renal brush border Na /H exchanger. Brush border membranes were solubilized with Triton X-100 and chromatographed sequentially over lentil lectin Sepharose 4B and 5-(A-benzyl-iV-ethyl)amiloride coupled to epoxy-activated Sepharose 6B. The eluant contained 178- and 146-kDa proteins that were susceptible to Endo-F. Moreover, the eluants reacted on dot blot immunoassays with antisera to a 20-amino acid peptide of a human Na /H exchanger vide infra). The relationship between these proteins and the 66-kDa protein previously identified by the same investigators using amiloride photolabeling is presently unclear. 

Wu, H. and Bruley, D.F, Homologous Human Blood Protein Separation Using Immobilized Metal Affinity Chromatography Protein C Separation from Prothrombin with Application to the Separation of Factor IX and Prothrombin, Biotechnol. Prog., 15, 928, 1999. 

Clemmit, R.H. and Chase, H.A., Immobilized metal affinity chromatography of P-galactosidase from unclarified Escherichia coli homogenates using expanded bed adsorption, /. Chromatogr. A, 874, 27, 2000. 

Brockman, A. H. Orlando, R. Probe immobilized affinity-chromatography mass-... 

Klenk, D.C., Hermanson, G.T., Krohn, R.I., Fujimoto, E.K., Mallia, A.K., Smith, P.K., England, J.D., Wiedmeyer, H.M., Little, R.R., and Goldstein, D.E. (1982) Determination of glycosylated hemoglobin by affinity chromatography Comparison with colorimetric and ion-exchange methods, and effects of common interferences. Clin. Chem. 28(10), 2088-2094. 

Purified C5-1 has been obtained from alfalfa leaf extracts by affinity chromatography on either a human IgG-Sepharose column or a Streamline rProtein A-Sepharose column. Interestingly, the purified product obtained with these two methods differed significantly. As shown in Fig. 1.5 a, the antibody fraction obtained from the human IgG column contained a mixture of different intermediate assembly forms of the heavy (H) and light (L) chains, ranging from H2 to the fully assembled H2L2 form. 

Figure 6.18 Preparative H PLC column (15 cm in diameter) used in processing of proteins required for therapeutic or diagnostic purposes. Column manufactured by Prochrom, Nancy, France (photograph courtesy of Affinity Chromatography Ltd)...

Hjelm H, Hjelm K, Sjoquist J. Protein A from Staphylococcus aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulins. FEBS Lett 1972 28 73-76. 

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