Immobilized metal affinity chromatography with fused silica microcolumns

Fused silica capillaries of 250 μm i.d. were slurry-packed with a TSK Chelate-5PW stationary phase. Cu(II) was immobilized on the iminodiacetate surface, and proteins were separated using either an increasing imidazole gradient or a decreasing pH gradient. The microcolumn system increases detection sensitivity without affecting chromatographic behavior. The possible use of this system in the study of tertiary structure relationships between related proteins is explored.

[1]  T. Yip,et al.  Protein interactions with immobilized transition metal ions: quantitative evaluations of variations in affinity and binding capacity. , 1990, Analytical biochemistry.

[2]  S. Snyder,et al.  Odorant-binding protein: odorant transport function in the vertebrate nasal epithelium , 1990 .

[3]  J. J. Edwards,et al.  Chromatographic separations of serum proteins on immobilized metal ion stationary phases. , 1989, Analytical biochemistry.

[4]  R. Tirindelli,et al.  Complete amino acid sequence of pyrazine-binding protein from cow nasal mucosa. , 1989, European journal of biochemistry.

[5]  M. Novotny,et al.  Fused silica packed microcolumns as micropreparative tools in protein analytical studies. , 1988, Analytical chemistry.

[6]  J. Porath,et al.  High-performance immobilized-metal-ion affinity chromatography of peptides and proteins. , 1988, Journal of chromatography.

[7]  M. Novotny,et al.  Separation of proteins by microcolumn liquid chromatography based on the reversed-phase and size-exclusion principles. , 1988, Journal of chromatography.

[8]  T. Yip,et al.  High-performance immobilized metal ion affinity chromatography of peptides: analytical separation of biologically active synthetic peptides. , 1988, Analytical biochemistry.

[9]  T. Yip,et al.  High-performance analytical applications of immobilized metal ion affinity chromatography. , 1987, Analytical biochemistry.

[10]  M. Novotny,et al.  High-efficiency microcolumn liquid chromatography separation and spectral characterization of nitrogen-containing polycyclics from fossil fuels , 1987 .

[11]  T. Yip,et al.  Interaction of proteins with immobilized Cu2+. Quantitation of adsorption capacity, adsorption isotherms and equilibrium constants by frontal analysis. , 1987, Journal of chromatography.

[12]  J. Porath,et al.  Selective adsorption of phosphoproteins on gel-immobilized ferric chelate. , 1986, Biochemistry.

[13]  E. Sulkowski.  Purification of proteins by IMAC , 1985 .

[14]  M. Novotny,et al.  Dead-volume free termination for packed columns in microcapillary liquid chromatography. , 1984, Analytical chemistry.

[15]  L. Andersson.  Fractionation of human serum proteins by immobilized metal affinity chromatography. , 1984, Journal of chromatography.

[16]  J. Porath,et al.  Immobilized metal ion affinity adsorption and immobilized metal ion affinity chromatography of biomaterials. Serum protein affinities for gel-immobilized iron and nickel ions. , 1983, Biochemistry.

[17]  H. Kikuchi,et al.  Significance of use of amino acids and histamine for the elution of nonhistone proteins in copper-chelate chromatography. , 1981, Analytical biochemistry.

[18]  J. Porath,et al.  Metal chelate affinity chromatography, a new approach to protein fractionation , 1975, Nature.