Skip to main content
SpringerLink
Log in

Printed glycan array: antibodies as probed in undiluted serum and effects of dilution

  • GlycoArray Section
  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

Using printed glycan array (PGA) we compared the results of antibody profiling in undiluted, moderately (1:15) and highly (1:100) diluted human blood serum. Undiluted serum is suitable for studying blood as a tissue in its native state, whereas to study the serum of newborns or small animals one usually has to dilute the starting material in order to have sufficient volume for PGA experimentation. The PGA used in this study allows for the use of whole serum without modifications to the protocol, and the background is surprisingly low. Antibodies profiles observed in undiluted serum versus 1:15 dilution were similar, with only a limited number of new signals identified in the undiluted serum. However, unexpected irregularities were found when IgG and IgM are measured separately, namely, at a 1:15 dilution more intensive IgG signals for many glycans are observed. We believe that in conditions of moderate dilution IgG and IgM antibodies can compete with each other for antigen and as a result, the higher affinity anti-glycan IgGs give rise to more intense signals. Therefore depending on the purpose, different dilutions of serum will be optimal: in competitive 1:15 conditions the observed IgG/IgM ratio corresponds to their titer, whereas at 1:100 dilution the measured ratio corresponds to real molar concentration of IgG and IgM.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Abbreviations

PGA:

Printed glycan array

Abs:

Antibodies

References

  1. Blixt, O., Head, S., Mondala, T., Scanlan, C., Huflejt, M.E., Alvarez, R., Bryan, M.C., Fazio, F., Calarese, D., Stevens, J., Razi, N., Stevens, D.J., Skehel, J.J., van Die, I., Burton, D.R., Wilson, I.A., Cummings, R., Bovin, N., Wong, C.-H., Paulson, J.C.: Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proc. Natl. Acad. Sci. USA 101, 17033–17038 (2004)

    Article  PubMed  CAS  Google Scholar 

  2. Huflejt, M.E., Vuskovic, M., Vasiliu, D., Xu, H., Obukhova, P., Shilova, N., Tuzikov, A., Galanina, O., Arun, B., Lu, K., Bovin, N.: Anti-carbohydrate antibodies of normal sera: findings, surprises and challenges. Mol. Immunol. 46, 3037–3049 (2009)

    Article  PubMed  CAS  Google Scholar 

  3. Obukhova, P., Korchagina, E., Henry, S., Bovin, N.: Natural anti-A and anti-B of the ABO system: allo- and auto-antibodies have different epitope specificity. Transfusion (Received for publication February 11, 2011; revision received July 11, 2011, and accepted August 16, 2011. doi:10.1111/j.1537-2995.2011.03381.x)

  4. Pochechueva, T., Jacob, F., Goldstein, D.R., Huflejt, M.E., Chinarev, A., Caduff, R., Fink, D., Hacker, N., Bovin, N.V., Heinzelmann-Schwarz, V.: Comparison of printed glycan array, suspension array and ELISA in the detection of human anti-glycan antibodies. Glycoconjugate J. 8–9, 507–517 (2011)

    Article  Google Scholar 

Download references

Acknowledgments

We would like to thank Drs. Galina Pazynina and Alexander Tuzikov (Institute of Bioorganic Chemistry, Moscow, Russia) for the synthesis of numerous glycans and constructive discussion; Kowa Chen for technical assistance with microarray printing. This study was supported in part by the RAS Presidium Program Molecular and Cell Biology, RFBR grant 10-04-01693 and the European Commission Marie Curie Program for support of the EuroGlycoArrays ITN, and Russian State Contract 16.512.11.2039 (code 2011-1.2-512-013-008).

Author information

Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    Nadezhda Shilova, Maxim Navakouski, Nailya Khasbiullina & Nicolai Bovin

  2. Copenhagen Center for Glycomics, Institute of Cellular and Molecular Medicine, Panum Institute, University of Copenhagen, Blegdamsvej 3, DK-2200, Copenhagen, Denmark

    Ola Blixt

Authors
  1. Nadezhda Shilova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maxim Navakouski
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nailya Khasbiullina
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ola Blixt
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nicolai Bovin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nicolai Bovin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shilova, N., Navakouski, M., Khasbiullina, N. et al. Printed glycan array: antibodies as probed in undiluted serum and effects of dilution. Glycoconj J 29, 87–91 (2012). https://doi.org/10.1007/s10719-011-9368-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10719-011-9368-8

Keywords

Search

Navigation