Controlled Pore Glass Chromatography Research Articles

Protein-sodium dodecyl sulfate complexes: Determination of molecular weight, size and shape by controlled pore glass chromatography

The peak position vs log molecular weight curves of protein-SDS complexes chromatographed on controlled pore glass of narrow pore size distribution is linear over a molecular weight range of 17,000–385,000. A glass with a pore size of approximately 500 Å allows the inclusion of all complexes in this range. Peak position curves on glasses with broad pore distributions show decreased resolution and deviate from linearity at low elution coefficients. Exclusion size analysis of the elution coefficients of individual complexes from different columns with pore diameters ranging from 197 to 650 Å gives from 120 to 423 Å as their longest dimension. Assuming constant hydration and SDS-to-protein ration, the found dimension suggests the shape of a football, rather than a sphere or rigid rod.

A single equation relating molecular weight, pore-size, and elution coefficient in the controlled pore glass chromatography of protein-sodium dodecyl sulfate complexes

A single equation relating molecular weight, pore-size, and elution coefficient in the controlled pore glass chromatography of protein-sodium dodecyl sulfate complexes

Virus isolation with glass of controlled pore size: MS2 bacteriophage and Kilham virus

Size-exclusion chromatography with controlled pore glass (CPG) was used in the further purification of renal brush border membrane vesicles (BBMV) isolated by the Ca precipitation method. The BBMV obtained had an almost spherical shape and their average diameter was about 95 nm in isotonic solution. The specific activities of alkaline phosphate and leucine aminopeptidase in the BBMV preparation were increased 18- and 17-fold, respectively, over those in the crude homogenate. The uptake of d-glucose by the purified BBMV in the presence of a sodium gradient reached 8.53 nmol/mg protein at 20 s. These results indicate that CPG chromatography is a suitable procedure by which to obtain purified renal BBMV of homogenous size and with high specific marker enzyme activity for use in the study of membrane transport.