Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011 May 15;83(10):3703-8.
doi: 10.1021/ac2001077. Epub 2011 Apr 19.

A comprehensive compositional analysis of heparin/heparan sulfate-derived disaccharides from human serum

Wei Wei  1 , Milady R Niñonuevo, Anish Sharma, Lieza M Danan-Leon, Julie A Leary
Affiliations

A comprehensive compositional analysis of heparin/heparan sulfate-derived disaccharides from human serum

Wei Wei et al. Anal Chem. .

Abstract

The analysis of heparan sulfate glycosaminoglycans (HSGAGs) variations in human serum at the disaccharide level has a great potential for disease diagnosis and prognosis. However, the lack of available analytical methodology for the compositional analysis of HSGAGs in human serum remains to be addressed to delineate the possible role of HSGAGs on the onset and/or progression of a disease. In this study, we have developed a method for the in-depth compositional analysis of the 12 heparin/HS-derived disaccharides from human serum using a combination of technologies--fractionation, exhaustive digestion, solid phase extraction, and LC-MS/MS. The method exhibits high recovery (72-110%) and good reproducibility (standard deviation of less than 5%) with a low limit of detection and quantification. Errors from the method validation were within 1.1%. Nondetectable non- or low-sulfated disaccharides in human serum were also detected using the optimized protocol. Further applying this method, the comprehensive analysis of HSGAGs compositions in human sera from female donors showed considerable variations in disaccharide patterns and compositions.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Comparison of MS flow injection and LC-MS analysis of a sample of six disaccharide standards (I-S, II-S, IV-A, II-H, II-A and internal standard I-P) in human serum matrix after SPE cleanup: (A) MS full scan spectrum shows a significant TFA dimer ion suppression; (B) LC-MS spectrum shows much better signals of disaccharides.
Figure 2
Figure 2
Comparison of the extracted ion chromatographs (XIC) from the MS1 and MS2 spectra: (A) XIC of I-S3- ([M-3H]3- at m/z 191.4) in MS1 spectrum (shown as an inset); (B) XIC of the diagnostic ion ([0,2A2]2- at m/z 217.9) of I-S in MS2 spectrum (shown as an inset); (C) XIC of I-A2- ([M-2H]2- at m/z 268.5) in MS1 spectrum. The inserted MS1 spectrum shows I-A ion and a matrix peak (m/z 269.9) that partially overlapped with I-A; (D) XIC of the diagnostic ion ([M-H2O-2H]2- at m/z 259.4) of I-A in MS2 spectrum (shown as an inset).
Figure 3
Figure 3
MS2 spectrum of isomeric disaccharides II-A/III-A (m/z 458.1 as the precursor ion) from the digested sample of human serum from platelet poor human plasma. The product ion m/z 357.0 is mostly contributed by [0,2A2]− fragment of II-A and m/z 236.9 corresponds to [B1]− fragment of III-A.
Figure 4
Figure 4
Bar graph of the compositions of the 12 heparin/HS-derived disaccharides from three single donor sera, 38 years old Caucasian female (Caucasian 38F), 44 years old African American female (African American 44F) and 29 years old Hispanic female (Hispanic 29F). The 12 disaccharides are shown in different colors as legend list below and the length of each color represents the percentage composition of the represented disaccharide. The number of sulfate groups in each 12 disaccharide is listed under the color legend accordingly.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

  • Biochemical signatures of disease severity in multiple sulfatase deficiency.
    Adang LA, Mowafy S, Herbst ZM, Zhou Z, Schlotawa L, Radhakrishnan K, Bentley B, Pham V, Yu E, Pillai NR, Orchard PJ, De Castro M, Vanderver A, Pasquali M, Gelb MH, Ahrens-Nicklas RC. Adang LA, et al. J Inherit Metab Dis. 2024 Mar;47(2):374-386. doi: 10.1002/jimd.12688. Epub 2023 Nov 1. J Inherit Metab Dis. 2024. PMID: 37870986
  • Noninvasive detection of any-stage cancer using free glycosaminoglycans.
    Bratulic S, Limeta A, Dabestani S, Birgisson H, Enblad G, Stålberg K, Hesselager G, Häggman M, Höglund M, Simonson OE, Stålberg P, Lindman H, Bång-Rudenstam A, Ekstrand M, Kumar G, Cavarretta I, Alfano M, Pellegrino F, Mandel-Clausen T, Salanti A, Maccari F, Galeotti F, Volpi N, Daugaard M, Belting M, Lundstam S, Stierner U, Nyman J, Bergman B, Edqvist PH, Levin M, Salonia A, Kjölhede H, Jonasch E, Nielsen J, Gatto F. Bratulic S, et al. Proc Natl Acad Sci U S A. 2022 Dec 13;119(50):e2115328119. doi: 10.1073/pnas.2115328119. Epub 2022 Dec 5. Proc Natl Acad Sci U S A. 2022. PMID: 36469776 Free PMC article.
  • Sulfated glycan recognition by carbohydrate sulfatases of the human gut microbiota.
    Luis AS, Baslé A, Byrne DP, Wright GSA, London JA, Jin C, Karlsson NG, Hansson GC, Eyers PA, Czjzek M, Barbeyron T, Yates EA, Martens EC, Cartmell A. Luis AS, et al. Nat Chem Biol. 2022 Aug;18(8):841-849. doi: 10.1038/s41589-022-01039-x. Epub 2022 Jun 16. Nat Chem Biol. 2022. PMID: 35710619 Free PMC article.
  • SARS-CoV-2: Receptor and Co-receptor Tropism Probability.
    Eslami N, Aghbash PS, Shamekh A, Entezari-Maleki T, Nahand JS, Sales AJ, Baghi HB. Eslami N, et al. Curr Microbiol. 2022 Mar 16;79(5):133. doi: 10.1007/s00284-022-02807-7. Curr Microbiol. 2022. PMID: 35292865 Free PMC article. Review.
  • Analysis of normal levels of free glycosaminoglycans in urine and plasma in adults.
    Bratulic S, Limeta A, Maccari F, Galeotti F, Volpi N, Levin M, Nielsen J, Gatto F. Bratulic S, et al. J Biol Chem. 2022 Feb;298(2):101575. doi: 10.1016/j.jbc.2022.101575. Epub 2022 Jan 8. J Biol Chem. 2022. PMID: 35007531 Free PMC article.
See all "Cited by" articles

References

    1. Saad OM, Leary JA. Anal. Chem. 2003;75:2985–2995. - PubMed
    1. Saad OM, Ebel H, Uchimura K, Rosen SD, Bertozzi CR, Leary JA. Glycobiology. 2005;15:818–826. - PubMed
    1. Meissen JK, Sweeney MD, Girardi M, Lawrence R, Esko JD, Leary JA. J. Am. Soc. Mass Spectrom. 2009;20:652–657. - PMC - PubMed
    1. Lawrence R, Lu H, Rosenberg RD, Esko JD, Zhang L. Nat. Methods. 2008;5:291–292. - PubMed
    1. Varki A, Cummings RD, Esko JD. Essentials of Glycobiology. 2nd ed. Vol. 16. Cold Spring Harbor Laboratory Press; New York: 2008. - PubMed

Publication types

-