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

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.

journal_name

Glycoconj J

journal_title

Glycoconjugate journal

authors

Shilova N,Navakouski M,Khasbiullina N,Blixt O,Bovin N

doi

10.1007/s10719-011-9368-8

subject

Has Abstract

pub_date

2012-04-01 00:00:00

pages

87-91

issue

2-3

eissn

0282-0080

issn

1573-4986

journal_volume

29

pub_type

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