Assay of isoenzymes by ion exchange chromatography

Details Assay of isoenzymes by ion exchange chromatography Assay of isoenzymes by ion exchange chromatography

1976-04-16

1977-09-06

Naff, David M.

Textiles: fiber preparation

23230B, 195 66R, 424 2, G01N 3104, G01N 3108, G01N 3114

Patent

active

040466345

ABSTRACT:
A simple, rapid anion-exchange column chromatographic technique is used to determine the presence of fractions of proteins and enzymes in human serum by separating the fractions on the basis of their net electrical charges. Samples of serum of unaltered ionic strength, layered on mini columns of an ion exchange agent previously equilibrated to a low ionic strength similar to blood serum with a buffer solution, are eluted stepwise with an appropriate buffer solution having different ionic strengths at each step. Elution of the samples is without previous dialysis or other ionization treatment. Column effluents are assayed for protein content or enzyme activity by conventional means, e.g., spectrophotometric analysis. Evaluation of sera from patients utilizing these methods reveals protein fractions or isoenzyme patterns which identify specific diseases associated with the protein or isoenzyme pattern.

REFERENCES:
patent: 3234199 (1966-02-01), Reid
Takahashi et al., Creative Phosphokinase Isozymes of Human Heart Muscle and Skeletal Muscle. Clinicz Chimicz Acta. vol. 38, 1972 (pp. 285-290).
Schmidt et al., An Improved Simple Chromatographic Method for Separating the Isoenzymes of Malic Dehydrigenase and Glutomic Oxaloacetic Transaminase. Clinicz Chimicz Acta. vol. 15, 1967, (pp. 337-342).
Richterich et al., A Study of Lactic Dehydrogenase Isoenzyme Pattern of Human Tissues by Adsorption-Elution on Sephadex-DEAE, Clinicz Chimicz Acta. vol. 8, 1963, (pp. 178-192).

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