Development of molecularly imprinted Acrylamide-Acrylamido phenylboronic acid copolymer microbeads for selective glycosaminoglycan separation in children urine

  • 1 Ege University, Faculty of Medicine, Medical Biochemistry Department, Bornova, 35100 İzmir, Turkey
  • 2 Ege University, Faculty of Science, Biochemistry Department, Bornova, İzmir, Turkey
  • 3 Ege University, Hospital of Children, Infant Nephrology Department, Bornova, İzmir, Turkey
  • 4 Ege University, Faculty of Medicine, Medical Biochemistry Department, Bornova, İzmir, Turkey
Zihni Onur UygunORCID iD: https://orcid.org/0000-0001-9045-7271, Burcu Okutucu, Şükriye Hacikara and Ferhan Sağın

Abstract

Background

In this study, we synthesized molecularly imprinted copolymers for liquid chromatography columns as a separator for glycosaminoglycan (dermatan sulfate; DS and chondroitin sulfate; CS) in urine.

Materials and methods

Acrylamide and acrylamido phenylboronic acid were used as monomers, acrylamide was used for as base monomer to attract negatively charged groups and acrylamido phenylboronic acid (AAPBA) residues used to form diol bonds between sugar and boronic acid residues to strengthen the attraction. These monomers were synthesized by using precipitation polymerization to form uniform spheres, which are more durable for the pressurized chromatographic systems. Trimethylolpropane trimethacrylate and AIBN were used as crosslinker and starter, respectively.

Results

These GAG selective polymers were filled by pressurized flow into the steel (4.6 mm × 1.6 mm) columns, then imprinted GAGs were extracted and analyzed to calculate binding capacity of each milligram polymer. Calibration curves of the GAG selective columns were obtained 62.5–1000 ng/mL less than 5% coefficient variation, and lower matrix effect.

Conclusion

Our imprinted columns separated different GAGs from urine specifically and sensitively. Matrix effect was at an ignorable level thus the challenging use.

    • Supplementary material
  • 1.

    Höök M, Kjellén L, Johansson S. Cell-surface glycosaminoglycans. Annu Rev Biochem 1984;53:847–69.

    • Crossref
    • PubMed
    • Export Citation
  • 2.

    Iozzo RV, Schaefer L. Proteoglycan form and function: a comprehensive nomenclature of proteoglycans. Matrix Biol 2015;42:11–55.

    • Crossref
    • PubMed
    • Export Citation
  • 3.

    Hitchcock AM, Yates KE, Shortkroff S, Costello CE, Zaia J. Optimized extraction of glycosaminoglycans from normal and osteoarthritic cartilage for glycomics profiling. Glycobiology 2007;17:25–35.

    • Crossref
    • PubMed
    • Export Citation
  • 4.

    Savolainen H. Isolation and separation of proteoglycans. J Chromatogr B Biomed Sci Appl 1999;722:255–62.

    • Crossref
    • PubMed
    • Export Citation
  • 5.

    Fuller M, Meikle PJ, Hopwood JJ. Glycosaminoglycan degradation fragments in mucopolysaccharidosis I. Glycobiology 2004;14:443–50.

    • Crossref
    • PubMed
    • Export Citation
  • 6.

    Byers S, Rozaklis T, Brumfield LK, Ranieri E, Hopwood JJ. Glycosaminoglycan accumulation and excretion in the mucopolysaccharidoses: characterization and basis of a diagnostic test for MPS. Mol Genet Metab 1998;65:282–90.

    • Crossref
    • PubMed
    • Export Citation
  • 7.

    Genovese F, Manresa AA, Leeming DJ, Karsdal MA, Boor P. The extracellular matrix in the kidney: a source of novel non-invasive biomarkers of kidney fibrosis? Fibrogenesis Tissue Repair 2014;7:4.

    • Crossref
    • PubMed
    • Export Citation
  • 8.

    Afratis N, Gialeli C, Nikitovic D, Tsegenidis T, Karousou E, Theocharis AD, et al. Glycosaminoglycans: key players in cancer cell biology and treatment. FEBS J 2012;279:1177–97.

    • Crossref
    • PubMed
    • Export Citation
  • 9.

    Kittlick PD. Inflammation, glycolytic metabolism, and glycosaminoglycans. Exp Pathol 1986;30:1–19.

    • Crossref
    • PubMed
    • Export Citation
  • 10.

    Page C, Rose M, Yacoub M, Pigott R. Antigenic heterogeneity of vascular endothelium. Am J Pathol 1992;141:673–83.

    • PubMed
    • Export Citation
  • 11.

    van Tiel J, Siebelt M, Reijman M, Bos PK, Waarsing JH, Zuurmond A-M, et al. Quantitative in vivo CT arthrography of the human osteoarthritic knee to estimate cartilage sulphated glycosaminoglycan content: correlation with ex-vivo reference standards. Osteoarthritis Cartilage 2016;24:1012–20.

    • Crossref
    • PubMed
    • Export Citation
  • 12.

    Wang JY, Roehrl MH. Glycosaminoglycans are a potential cause of rheumatoid arthritis. Proc Natl Acad Sci USA 2002;99: 14362–7.

    • Crossref
    • Export Citation
  • 13.

    Millington D, Zhang H, Young S, Orchard P, Tolar J, Auray-Blais C. Analysis of glycosaminoglycans in cerebrospinal fluid using tandem mass spectrometry: potential for therapeutic monitoring of patients with mucopolysaccharidoses. Mol Genet Metab 2011;102:S29–30.

    • Crossref
    • Export Citation
  • 14.

    Yang B, Chang Y, Weyers AM, Sterner E, Linhardt RJ. Disaccharide analysis of glycosaminoglycan mixtures by ultra-high-performance liquid chromatography-mass spectrometry. J Chromatogr A 2012;1225:91–8.

    • Crossref
    • PubMed
    • Export Citation
  • 15.

    Whitley CB, Ridnour MD, Draper KA, Dutton CM, Neglia JP. Diagnostic test for mucopolysaccharidosis. I. Direct method for quantifying excessive urinary glycosaminoglycan excretion. Clin Chem 1989;35:374–9.

    • Crossref
    • PubMed
    • Export Citation
  • 16.

    Terho TT, Hartiala K. Method for determination of the sulfate content of glycosaminoglycans. Anal Biochem 1971;41:471–6.

    • Crossref
    • PubMed
    • Export Citation
  • 17.

    Berry HK. Screening for mucopolysaccharide disorders with the Berry spot test. Clin Biochem 1987;20:365–71.

    • Crossref
    • PubMed
    • Export Citation
  • 18.

    Maccari F, Volpi N. Glycosaminoglycan blotting on nitrocellulose membranes treated with cetylpyridinium chloride after agarose-gel electrophoretic separation. Electrophoresis 2002;23:3270–7.

    • Crossref
    • PubMed
    • Export Citation
  • 19.

    Coutinho MF, Lacerda L, Alves S. Glycosaminoglycan storage disorders: a review. Biochem Res Int 2012;2012:471325.

    • PubMed
    • Export Citation
  • 20.

    Komosinska-Vassev K, Blat D, Olczyk P, Szeremeta A, Jura-Półtorak A, Winsz-Szczotka K, et al. Urinary glycosaminoglycan (uGAG) excretion in healthy pediatric and adolescent population. Clin Biochem 2014;47:1341–3.

    • Crossref
    • PubMed
    • Export Citation
  • 21.

    Wang X-H, Xie L-F, Dong Q, Liu H-L, Huang Y-P, Liu Z-S. Synthesis of monodisperse molecularly imprinted microspheres with multi-recognition ability via precipitation polymerization for the selective extraction of cyromazine, melamine, triamterene and trimethoprim. J Chromatogr B Analyt Technol Biomed Life Sci 2015;1007:127–31.

    • Crossref
    • PubMed
    • Export Citation
  • 22.

    Springsteen G, Wang B. A detailed examination of boronic acid-diol complexation. Tetrahedron 2002;58:5291–300.

    • Crossref
    • Export Citation
  • 23.

    Ye L, Weiss R, Mosbach K. Synthesis and characterization of molecularly imprinted microspheres. Macromolecules 2000;33:8239–45.

    • Crossref
    • Export Citation
  • 24.

    Okutucu B, Önal S. Molecularly imprinted polymers for separation of various sugars from human urine. Talanta 2011;87:74–9.

    • Crossref
    • PubMed
    • Export Citation
Purchase article
Get instant unlimited access to the article.
$42.00
Log in
Already have access? Please log in.


or
Log in with your institution

Journal + Issues

Turkish Journal of Biochemistry (TJB), official journal of Turkish Biochemical Society, is issued electronically every 2 months. The main aim of the journal is to support the research and publishing culture by ensuring that every published manuscript has an added value and thus providing international acceptance of the “readability” of the manuscripts published in the journal.

Search