Skip to main content
SpringerLink
Log in

Purification of Capsular Polysaccharide Complex from Gram-Negative Bacteria

  • Protocol
  • First Online:
Bacterial Polysaccharides

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1954))

Abstract

Capsular polysaccharides are a dominant class of antigens from bacteria, both pathogenic and symbiotic or commensal. With the rise of awareness for the influence of the microbiota over immune system development and immune homeostasis, analysis of the antigens is more important than ever. Here we describe a method for the isolation of capsular polysaccharide from gram-negative bacteria, with the purification of polysaccharide from the commensal bacterium Bacteroides fragilis serving as an example. The method efficiently removes all detectable endotoxins and other lipid components, proteins, and nucleic acids, providing a source of capsular polysaccharide for immunologic study.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Falk PG, Hooper LV, Midtvedt T et al (1998) Creating and maintaining the gastrointestinal ecosystem: what we know and need to know from gnotobiology. Microbiol Mol Biol Rev 62:1157–1170

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Hooper LV, Midtvedt T, Gordon JI (2002) How host-microbial interactions shape the nutrient environment of the mammalian intestine. Annu Rev Nutr 22:283–307

    Article  CAS  Google Scholar 

  3. Nishio J, Honda K (2012) Immunoregulation by the gut microbiota. Cell Mol Life Sci 69:3635–3650

    Article  CAS  Google Scholar 

  4. Peterson CT, Sharma V, Elmen L et al (2015) Immune homeostasis, dysbiosis and therapeutic modulation of the gut microbiota. Clin Exp Immunol 179:363–377

    Article  CAS  Google Scholar 

  5. Johnson JL, Jones MB, Cobb BA (2015) Bacterial capsular polysaccharide prevents the onset of asthma through T-cell activation. Glycobiology 25:368–375

    Article  CAS  Google Scholar 

  6. Johnson JL, Jones MB, Cobb BA (2018) Polysaccharide-experienced effector T cells induce IL-10 in FoxP3+ regulatory T cells to prevent pulmonary inflammation. Glycobiology 28:50–58

    Article  CAS  Google Scholar 

  7. Mazmanian SK, Round JL, Kasper DL (2008) A microbial symbiosis factor prevents intestinal inflammatory disease. Nature 453(7195):620–625

    Article  CAS  Google Scholar 

  8. Ochoa-Reparaz J, Mielcarz DW, Wang Y et al (2010) A polysaccharide from the human commensal Bacteroides fragilis protects against CNS demyelinating disease. Mucosal Immunol 3:487–495

    Article  CAS  Google Scholar 

  9. Tzianabos AO, Onderdonk AB, Rosner B et al (1993) Structural features of polysaccharides that induce intra-abdominal abscesses. Science 262(5132):416–419

    Article  CAS  Google Scholar 

  10. Krinos CM, Coyne MJ, Weinacht KG et al (2001) Extensive surface diversity of a commensal microorganism by multiple DNA inversions. Nature 414(6863):555–558

    Article  CAS  Google Scholar 

  11. Tzianabos AO, Pantosti A, Baumann H et al (1992) The capsular polysaccharide of Bacteroides fragilis comprises two ionically linked polysaccharides. J Biol Chem 267:18230–18235

    CAS  PubMed  Google Scholar 

  12. Cobb BA, Wang Q, Tzianabos AO et al (2004) Polysaccharide processing and presentation by the MHCII pathway. Cell 117:677–687

    Article  CAS  Google Scholar 

  13. Johnson JL, Jones MB, Cobb BA (2015) Polysaccharide A from the capsule of Bacteroides fragilis induces clonal CD4+ T cell expansion. J Biol Chem 290:5007–5014

    Article  CAS  Google Scholar 

  14. Tzianabos AO, Pantosti A, Baumann H et al (1991) Structural characterization of two surface polysaccharides of Bacteroides fragilis. Trans Assoc Am Phys 104:285–295

    CAS  PubMed  Google Scholar 

  15. Kreisman LS, Friedman JH, Neaga A et al (2007) Structure and function relations with a T-cell-activating polysaccharide antigen using circular dichroism. Glycobiology 17:46–55

    Article  CAS  Google Scholar 

  16. Cobb BA, Kasper DL (2008) Characteristics of carbohydrate antigen binding to the presentation protein HLA-DR. Glycobiology 18:707–718

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by NIH grants GM115234 and GM082916 to BAC, and AI089474 to CAA.

Author information

Authors and Affiliations

  1. Case Western Reserve University School of Medicine, Cleveland, OH, USA

    Carlos A. Alvarez & Brian A. Cobb

Authors
  1. Carlos A. Alvarez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Brian A. Cobb
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Brian A. Cobb .

Editor information

Editors and Affiliations

  1. Department of Biomedical and Molecular Sciences, Queen’s University, Kingston, ON, Canada

    Inka Brockhausen

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Alvarez, C.A., Cobb, B.A. (2019). Purification of Capsular Polysaccharide Complex from Gram-Negative Bacteria. In: Brockhausen, I. (eds) Bacterial Polysaccharides. Methods in Molecular Biology, vol 1954. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9154-9_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9154-9_3

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9153-2

  • Online ISBN: 978-1-4939-9154-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation