Development and evaluation of a fluorescence microplate assay for quantification of heparins and other sulfated carbohydrates

Lühn, Susanne; Schräder, Thomas; Sun, Wei; Alban, Susanne

doi:10.1016/j.jpba.2009.12.014

Cited by 20 publications

(21 citation statements)

References 29 publications

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“…The linear range only slightly differed from that of heparin (0.3 to 5.0 µg/mL) (Figure 2 and [25]). According to the known degree of sulfation dependence the slope of the line was smaller and the maximum FI increase was lower due to the lower degrees of sulfation of D.s .-SP indicating weaker electrostatic interactions with the ethylammonium moieties of Polymer-H [25,26]. Interestingly, the effect of D.s.…”

Section: Resultsmentioning

confidence: 99%

“…It proved to detect a semi-synthetic linear homoglucan sulfate with a degree of sulfation as low as 0.60, but not another one with a still lower degree of sulfation of 0.25 [25]. The FI increase of Polymer-H by the sulfated homoglucan with degree of sulfation of 0.60 was about four times lower than that by heparin and a homoglucan sulfate with a degree of sulfation of 1.26.…”

Section: Resultsmentioning

confidence: 99%

“…Thus, the branched sulfated xylogalactan chains of D.s. -SP seem to interact with the aminomethylphenylboronic acid units of Polymer-H similar to the sulfated glycosaminoglycan chains of heparin, maybe by forming boronic esters [25,27]. The linear range only slightly differed from that of heparin (0.3 to 5.0 µg/mL) (Figure 2 and [25]).…”

Section: Resultsmentioning

confidence: 99%

“…-SP seem to interact with the aminomethylphenylboronic acid units of Polymer-H similar to the sulfated glycosaminoglycan chains of heparin, maybe by forming boronic esters [25,27]. The linear range only slightly differed from that of heparin (0.3 to 5.0 µg/mL) (Figure 2 and [25]). According to the known degree of sulfation dependence the slope of the line was smaller and the maximum FI increase was lower due to the lower degrees of sulfation of D.s .-SP indicating weaker electrostatic interactions with the ethylammonium moieties of Polymer-H [25,26].…”

Section: Resultsmentioning

confidence: 99%

“…Recently, we developed a sensitive microplate assay for quantification of heparins, which is based on the fluorescence intensity (FI) increase of the heparin sensor Polymer-H [25]. Polymer-H consists of a methacrylamide skeleton linked with three functional units: (1) ortho -aminomethylphenylboronic acid units forming boronic esters with glycans; (2) ethylammonium moieties electrostatically binding to anionic sulfated glycans; and (3) fluorescent dansyl monomers [26].…”

Section: Introductionmentioning

confidence: 99%

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Simple and Rapid Quality Control of Sulfated Glycans by a Fluorescence Sensor Assay—Exemplarily Developed for the Sulfated Polysaccharides from Red Algae Delesseria sanguinea

2014

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Sulfated polysaccharides (SP) from algae are of great interest due to their manifold biological activities. Obstacles to commercial (especially medical) application include considerable variability and complex chemical composition making the analysis and the quality control challenging. The aim of this study was to evaluate a simple microplate assay for screening the quality of SP. It is based on the fluorescence intensity (FI) increase of the sensor molecule Polymer-H by SP and was originally developed for direct quantification of SP. Exemplarily, 65 SP batches isolated from the red alga Delesseria sanguinea (D.s.-SP) and several other algae polysaccharides were investigated. Their FI increase in the Polymer-H assay was compared with other analytical parameters. By testing just one concentration of a D.s.-SP sample, quality deviations from the reference D.s.-SP and thus both batch-to-batch variability and stability can be detected. Further, structurally distinct SP showed to differ in their concentration-dependent FI profiles. By using corresponding reference compounds, the Polymer-H assay is therefore applicable as identification assay with high negative predictability. In conclusion, the Polymer-H assay showed to represent not only a simple method for quantification, but also for characterization identification and differentiation of SP of marine origin.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Simple and Rapid Quality Control of Sulfated Glycans by a Fluorescence Sensor Assay—Exemplarily Developed for the Sulfated Polysaccharides from Red Algae Delesseria sanguinea

2014

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Proteoglycans and Acidic Polysaccharides Analysis

Lauder

2015

Encyclopedia of Analytical Chemistry

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The study of the structure and function of proteoglycans (PGs) and glycosaminoglycans (GAGs) has assumed an increasing significance as the wide range of important biological processes in which they play central roles has been elucidated. As studies of GAG structure expand in scope, and detail, the full structural diversity of these polymers is being uncovered. Significant heterogeneity can be seen but relationships with development, age, and pathology are being uncovered, which place these macromolecules centrally in these processes. Available methodologies allow detailed analysis of GAGs, but careful attention to the sample requirements of these methods and to their limitations is required to ensure the quality and reliability of the data derived is assured. Important developments, including mass spectrometry approaches, will take the field forward but well‐established methods for the isolation and characterization of these complex molecules continue to have a hugely important role to play in structural and functional studies. This article examines a wide range of methodologies for the isolation and characterization of the PGs and GAGs in which recent developments in the field are reported along with well‐established methods for the characterization of entire GAG chains or discrete features within those.

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Low-Molecular-Weight Heparins: Differential Characterization/Physical Characterization

Guerrini

Bisio

2011

Handbook of Experimental Pharmacology

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Low-molecular-weight heparins (LMWHs), derived from unfractionated heparin (UFH) through different depolymerization processes, have advantages with respect to the parent heparin in terms of pharmacokinetics, convenience of administration, and reduced side effects. Each LMWH can be considered as an independent drug with its own activity profile, placing significance on their biophysical characterization, which will also enable a better understanding of their structure-function relationship. Several chemical and physical methods, some involving sample modification, are now available and are reviewed.

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Development and evaluation of a fluorescence microplate assay for quantification of heparins and other sulfated carbohydrates

Cited by 20 publications

References 29 publications

Simple and Rapid Quality Control of Sulfated Glycans by a Fluorescence Sensor Assay—Exemplarily Developed for the Sulfated Polysaccharides from Red Algae Delesseria sanguinea

Simple and Rapid Quality Control of Sulfated Glycans by a Fluorescence Sensor Assay—Exemplarily Developed for the Sulfated Polysaccharides from Red Algae Delesseria sanguinea

Proteoglycans and Acidic Polysaccharides Analysis

Low-Molecular-Weight Heparins: Differential Characterization/Physical Characterization

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