Quantitation of proteoglycans as glycosaminoglycans in biological fluids using an alcian blue dot blot analysis - PubMed (original) (raw)
. 1998 Feb 15;256(2):229-37.
doi: 10.1006/abio.1997.2494.
Affiliations
- PMID: 9473282
- DOI: 10.1006/abio.1997.2494
Quantitation of proteoglycans as glycosaminoglycans in biological fluids using an alcian blue dot blot analysis
S Björnsson. Anal Biochem. 1998.
Abstract
A method for quantitation of intact proteoglycans as GAGs in biological fluids (blood plasma, synovial fluid) or 4 M guanidine extracts of tissues has been published previously (S. Björnsson, Anal. Biochem. 210, 282-291, 1993). The method is based on the specific interaction between sulfated polymers and the tetravalent cationic dye Alcian blue at pH 1.5 in 0.4 M guanidine-HCl and in the presence of 0.25% Triton. The absorbance assay has a measuring range of 1-20 microgram of glycosaminoglycan (GAG) which is not sensitive enough to measure the low contents of proteoglycans in blood plasma, urine, or wound fluid. A dot blot assay is now described in which the Alcian blue-GAG complexes are collected on a polyvinylidene fluoride membrane, by filtration in a dot blot apparatus, and the stain is quantitated as reflectance by scanning and densitometry. The assay requires 10 microliter of sample and has a measuring range of 10-800 ng of GAG, corresponding to a concentration of 1-80 mg/liter, suitable for proteoglycans in biological fluids. The procedures for chemistry, scanning, densitometry, and curve fitting were each evaluated separately. The error contributed by chemistry accounted for a minor portion of the imprecision. The imprecision contributed by scanning was the most important source of within-run and between-run imprecision, and was caused by inequalities of the charge-coupled device along the scanning arm. Unexpectedly, curve fitting was also a major source of total imprecision in dot blot quantitation and differed with the type of equation used. The between-run imprecision calculated as CV (SD/mean . 100) was 13.0% at 8 mg/liter. The response of the assay was identical for six different commercial preparations of GAGs (chondroitin-4-sulfate, chondroitin-6-sulfate, dermatan sulfate, keratan sulfate, heparan sulfate, and heparin) despite differences in degree of sulfation known to exist. There was no positive or negative interference by blood plasma, apart from a slight negative interference on the quantitation of heparan sulfate. Analysis of 319 paired blood plasma and urine specimens from hospitalized patients showed a variation of plasma GAGs of 0.1-17.6 and urine-GAGs of 0.0-45.6 mg/liter. There was no correlation between plasma and urine GAG concentrations.
Copyright 1998 Academic Press.
Similar articles
- Simultaneous preparation and quantitation of proteoglycans by precipitation with alcian blue.
Björnsson S. Björnsson S. Anal Biochem. 1993 May 1;210(2):282-91. doi: 10.1006/abio.1993.1197. Anal Biochem. 1993. PMID: 8512063 - Binding and detection of glycosaminoglycans immobilized on membranes treated with cationic detergents.
Karlsson M, Edfors-Lilja I, Björnsson S. Karlsson M, et al. Anal Biochem. 2000 Nov 1;286(1):51-8. doi: 10.1006/abio.2000.4767. Anal Biochem. 2000. PMID: 11038273 - Electrophoresis and detection of nanogram quantities of exogenous and endogenous glycosaminoglycans in biological fluids.
al-Hakim A, Linhardt RJ. al-Hakim A, et al. Appl Theor Electrophor. 1991;1(6):305-12. Appl Theor Electrophor. 1991. PMID: 1718441 - Analysis of the Glycosaminoglycan Chains of Proteoglycans.
Song Y, Zhang F, Linhardt RJ. Song Y, et al. J Histochem Cytochem. 2021 Feb;69(2):121-135. doi: 10.1369/0022155420937154. Epub 2020 Jul 6. J Histochem Cytochem. 2021. PMID: 32623943 Free PMC article. Review. - Disease-specific glycosaminoglycan patterns in the extracellular matrix of human lung and brain.
Ennemoser M, Pum A, Kungl A. Ennemoser M, et al. Carbohydr Res. 2022 Jan;511:108480. doi: 10.1016/j.carres.2021.108480. Epub 2021 Nov 17. Carbohydr Res. 2022. PMID: 34837849 Review.
Cited by
- Endogenous Glycosaminoglycans in Various Pathologic Plasma Samples as Measured by a Fluorescent Quenching Method.
Kantarcioglu B, Mehrotra S, Papineni C, Siddiqui F, Kouta A, Hoppensteadt D, Bansal V, Darki A, Van Thiel DH, Fareed J. Kantarcioglu B, et al. Clin Appl Thromb Hemost. 2022 Jan-Dec;28:10760296221144047. doi: 10.1177/10760296221144047. Clin Appl Thromb Hemost. 2022. PMID: 36474353 Free PMC article. - Abundance and size of hyaluronan in naked mole-rat tissues and plasma.
Del Marmol D, Holtze S, Kichler N, Sahm A, Bihin B, Bourguignon V, Dogné S, Szafranski K, Hildebrandt TB, Flamion B. Del Marmol D, et al. Sci Rep. 2021 Apr 12;11(1):7951. doi: 10.1038/s41598-021-86967-9. Sci Rep. 2021. PMID: 33846452 Free PMC article. - Newborn screening of mucopolysaccharidoses: past, present, and future.
Arunkumar N, Langan TJ, Stapleton M, Kubaski F, Mason RW, Singh R, Kobayashi H, Yamaguchi S, Suzuki Y, Orii K, Orii T, Fukao T, Tomatsu S. Arunkumar N, et al. J Hum Genet. 2020 Jul;65(7):557-567. doi: 10.1038/s10038-020-0744-8. Epub 2020 Apr 10. J Hum Genet. 2020. PMID: 32277174 Review. - Advances in glycosaminoglycan detection.
Khan SA, Mason RW, Kobayashi H, Yamaguchi S, Tomatsu S. Khan SA, et al. Mol Genet Metab. 2020 Jun;130(2):101-109. doi: 10.1016/j.ymgme.2020.03.004. Epub 2020 Mar 27. Mol Genet Metab. 2020. PMID: 32247585 Free PMC article. Review. - Integrated Multi-Assay Culture Model for Stem Cell Chondrogenic Differentiation.
Prosser A, Scotchford C, Roberts G, Grant D, Sottile V. Prosser A, et al. Int J Mol Sci. 2019 Feb 22;20(4):951. doi: 10.3390/ijms20040951. Int J Mol Sci. 2019. PMID: 30813231 Free PMC article.