Quantification of alginate by aggregation induced by calcium ions and fluorescent polycations

Research output: Research - peer-reviewArticle

  • 4 Citations

Abstract

For quantification of polysaccharides, including heparins and alginates, the commonly used carbazole assay involves hydrolysis of the polysaccharide to form a mixture of UV-active dye conjugate products. Here, we describe two efficient detection and quantification methods that make use of the negative charges of the alginate polymer and do not involve degradation of the targeted polysaccharide. The first method utilizes calcium ions to induce formation of hydrogel-like aggregates with alginate polymer; the aggregates can be quantified readily by staining with a crystal violet dye. This method does not require purification of alginate from the culture medium and can measure the large amount of alginate that is produced by a mucoid Pseudomonas aeruginosa culture. The second method employs polycations tethering a fluorescent dye to form suspension aggregates with the alginate polyanion. Encasing the fluorescent dye in the aggregates provides an increased scattering intensity with a sensitivity comparable to that of the conventional carbazole assay. Both approaches provide efficient methods for monitoring alginate production by mucoid P. aeruginosa.

LanguageEnglish (US)
Pages76-81
Number of pages6
JournalAnalytical Biochemistry
Volume492
DOIs
StatePublished - Jan 1 2016

Fingerprint

Agglomeration
Ions
Calcium
alginic acid
polycations
Polysaccharides
Fluorescent Dyes
Assays
Polymers
Coloring Agents
carbazole
Pseudomonas aeruginosa
Alginates
Gentian Violet
Hydrogel
Purification
Culture Media
Heparin
Hydrolysis
Suspensions

Keywords

  • Aggregation
  • Alginate
  • Increased scattering
  • Mucoid Pseudomonas aeruginosa
  • Multivalent binding

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

Cite this

Quantification of alginate by aggregation induced by calcium ions and fluorescent polycations. / Zheng, Hewen; Korendovych, Ivan V.; Luk, Yan Yeung.

In: Analytical Biochemistry, Vol. 492, 01.01.2016, p. 76-81.

Research output: Research - peer-reviewArticle

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