Skip to main content
SpringerLink
Log in
Book cover

Electrophoretic Separation of Proteins pp 387–401Cite as

Isoelectric Focusing in Agarose Gel for Detection of Oligoclonal Bands in Cerebrospinal and Other Biological Fluids

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1855))

Abstract

Isoelectric focusing (IEF) coupled with immunodetection (immunofixation or immunoblotting) has become the leading technique for the detection and study of oligoclonal bands (OCBs) in cerebrospinal fluid (CSF) and also is increasingly used in other body fluids such as the tear and serum. Limited commercial availability of precast agarose IEF gels for research and a need for customization prompted reporting a detailed general protocol for the preparation and casting of agarose IEF gel along with sample, control, and isoelectric point marker preparation and carrying out the focusing itself for CSF OCBs. However, the method is readily adaptable to the use of other body fluid specimens and, possibly, research specimens such as culture fluids as well.

This is a preview of subscription content, 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   189.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD   249.99
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Polman CH, Reingold SC, Edan G et al (2005) Diagnostic criteria for multiple sclerosis: 2005 revisions to the “McDonald Criteria”. Ann Neurol 58:840–846

    Article  Google Scholar 

  2. Freedman MS, Thompson EJ, Deisenhammer F et al (2005) Recommended standard of cerebrospinal fluid analysis in the diagnosis of multiple sclerosis: a consensus statement. Arch Neurol 62:865–870

    Article  Google Scholar 

  3. Link H, Huang Y-M (2006) Oligoclonal bands in multiple sclerosis cerebrospinal fluid: An update on methodology and clinical usefulness. J Neuroimmunol 180:17–28

    Article  CAS  Google Scholar 

  4. Regeniter A, Kuhle J, Mehling M et al (2009) A modern approach to CSF analysis: pathophysiology, clinical application, proof of concept and laboratory reporting. Clin Neurol Neurosurg 111:313–318

    Article  Google Scholar 

  5. Schäffler N, Köpke S, Winkler L et al (2011) Accuracy of diagnostic tests in multiple sclerosis—a systematic review. Acta Neurol Scand 124:151–164

    Article  Google Scholar 

  6. Devos D, Forzy G, de Seze J et al (2001) Silver stained isoelectrophoresis of tears and cerebrospinal fluid in multiple sclerosis. J Neurol 248:672–675

    Article  CAS  Google Scholar 

  7. Calais G, Forzy G, Crinquette C et al (2010) Tear analysis in clinically isolated syndrome as new multiple sclerosis criterion. Mult Scler 16:87–92

    Article  Google Scholar 

  8. Fortini AS, Sanders EL, Weinshenker BG et al (2003) Cerebrospinal fluid oligoclonal bands in the diagnosis of multiple sclerosis. Isoelectric focusing with IgG immunoblotting compared with high-resolution agarose gel electrophoresis and cerebrospinal fluid IgG index. Am J Clin Pathol 120:672–675

    Article  CAS  Google Scholar 

  9. Nakashima I, Fujihara K, Sato S et al (2005) Oligoclonal IgG bands in Japanese patients with multiple sclerosis. A comparative study between isoelectric focusing with IgG immunofixation and high-resolution agarose gel electrophoresis. J Neuroimmunol 159:133–136

    Article  CAS  Google Scholar 

  10. Sanders E, Katzmann JA, Clark R et al (1999) Development of capillary electrophore.is as an alternative to high resolution agarose electrophoresis for the diagnosis of multiple sclerosis. Clin Chem Lab Med 37:37–45

    CAS  PubMed  Google Scholar 

  11. Barnidge DR, Kohlhagen MC, Zheng S et al (2015) Monitoring oligoclonal immunoglobulins in cerebral spinal fluid using microLC-ESI-Q-TOF mass spectrometry. J Neuroimmunol 288:123–126

    Article  CAS  Google Scholar 

  12. Laurenzi MA, Link H (1979) Characterisation of the mobility on isoelectric focusing of individual proteins in CSF and serum by immunofixation. J Neurol Neurosurg Psychiatry 42:368–372

    Article  CAS  Google Scholar 

  13. Mehta PD (1991) Diagnostic usefulness of cerebrospinal fluid in multiple sclerosis. Crit Rev Clin Lab Sci 28:233–251

    Article  CAS  Google Scholar 

  14. Sharief MK, Thompson EJ (1992) Distribution of cerebrospinal fluid oligoclonal IgM bands in neurological diseases: a comparison between agarose electrophoresis and isoelectric focusing. J Neurol Sci 109:83–87

    Article  CAS  Google Scholar 

  15. Mehta PD, Patrick BA, Thormar H et al (1982) Oligoclonal IgG bands with and without measles antibody activity in sera of patients with subacute sclerosing panencephalitis (SSPE). J Immunol 129:1983–1985

    CAS  PubMed  Google Scholar 

  16. Virtanen JO, Wohler J, Fenton K et al (2014) Oligoclonal bands in multiple sclerosis reactive against two herpesviruses and association with magnetic resonance imaging findings. Mult Scler 20:27–34

    Article  CAS  Google Scholar 

  17. Storstein A, Monstad SE, Honnorat J et al (2004) Paraneoplastic antibodies detected by isoelectric focusing of cerebrospinal fluid and serum. Neuroimmunol 155:150–154

    Article  CAS  Google Scholar 

  18. Stich O, Rauer S (2007) Antigen-specific oligoclonal bands in cerebrospinal fluid and serum from patients with anti-amphiphysin- and anti-CV2/CRMP5 associated paraneoplastic neurological syndromes. Eur J Neurol 14:650–653

    Article  CAS  Google Scholar 

  19. Franciotta D, Lolli F (2007) Interlaboratory reproducibility of isoelectric focusing in oligoclonal band detection. [Letter]. Clin Chem 53:1557–1558

    Article  CAS  Google Scholar 

  20. Lin MW, Suan D, Lenton K et al (2012) Differentiating patterns of oligoclonal banding in the cerebrospinal fluid improves diagnostic utility for multiple sclerosis. Pathology 44:248–250

    Article  Google Scholar 

  21. Boudet S, Peyrodie L, Wang Z et al (2016) Semi-automated image analysis of gel electrophoresis of cerebrospinal fluid for oligoclonal band detection. Conf Proc IEEE Eng Med Biol Soc 2016:744–747

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, MD, USA

    Gyorgy Csako

Authors
  1. Gyorgy Csako
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Biji T. Kurien

  2. Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    R. Hal Scofield

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

Csako, G. (2019). Isoelectric Focusing in Agarose Gel for Detection of Oligoclonal Bands in Cerebrospinal and Other Biological Fluids. In: Kurien, B., Scofield, R. (eds) Electrophoretic Separation of Proteins. Methods in Molecular Biology, vol 1855. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8793-1_33

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8793-1_33

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8792-4

  • Online ISBN: 978-1-4939-8793-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation