Isolation and purification of proteoglycans

Research output: Contribution to journalArticle

Abstract

Purification of a protein typically involves development of a quantitative assay to track protein integrity (e.g. enzyme activity) during subsequent isolation steps. The generalized procedure involves choosing the source of the protein, defining extraction conditions, developing bulk purification methods followed by refined, more selective methods. The purification of proteoglycans is often complicated by a) limited source quantities, b) necessity of chaotropic solvents for efficient extraction, c) their large molecular size and d) lack of defined functions to enable purity (i.e. activity, conformation) to be assessed. Because the usual goal of proteoglycan purification is physical characterization (intact molecular weight, core protein and glycosaminoglycan class and size), the problems of a suitable assay and/or native conformation are avoided. The 'assay' for tracking proteoglycan isolation typically utilizes uronic acid content or radiolabel incorporation as a marker. Once extracted from their cellular/extracellular environment, proteoglycans can be isolated by density gradient centrifugation and/or column chromatography techniques. Recent advances in the composition of chromatographic supports have enabled the application of ion-exchange, gel permeation, hydrophobic interaction and affinity chromatography resins using efficient high-pressure liquid chromatography to proteoglycan purification.

Original languageEnglish (US)
Pages (from-to)369-383
Number of pages15
JournalExperientia
Volume49
Issue number5
DOIs
StatePublished - May 1993
Externally publishedYes

Fingerprint

Proteoglycans
Purification
Assays
Conformations
Proteins
High pressure liquid chromatography
Uronic Acids
Affinity chromatography
Column chromatography
Density Gradient Centrifugation
Centrifugation
Ion Exchange
Enzyme activity
Glycosaminoglycans
Hydrophobic and Hydrophilic Interactions
Affinity Chromatography
Permeation
Chromatography
Ion exchange
Resins

Keywords

  • assays
  • chromatography
  • extraction
  • Proteoglycans
  • purification
  • ultracentrifugation

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Isolation and purification of proteoglycans. / Fedarko, Neal S.

In: Experientia, Vol. 49, No. 5, 05.1993, p. 369-383.

Research output: Contribution to journalArticle

Fedarko, Neal S. / Isolation and purification of proteoglycans. In: Experientia. 1993 ; Vol. 49, No. 5. pp. 369-383.
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