An optimized enzyme-linked lectin assay to measure influenza A virus neuraminidase inhibition antibody titers in human sera

Laura Couzens; Jin Gao; Kim Westgeest; Matthew Sandbulte; Vladimir Lugovtsev; Ron Fouchier; Maryna Eichelberger

doi:10.1016/j.jviromet.2014.09.003

An optimized enzyme-linked lectin assay to measure influenza A virus neuraminidase inhibition antibody titers in human sera

J Virol Methods. 2014 Dec 15:210:7-14. doi: 10.1016/j.jviromet.2014.09.003. Epub 2014 Sep 16.

Authors

Laura Couzens¹, Jin Gao², Kim Westgeest³, Matthew Sandbulte⁴, Vladimir Lugovtsev⁵, Ron Fouchier⁶, Maryna Eichelberger⁷

Affiliations

¹ Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Avenue, Building 72, HFM445, Silver Spring, MD 20993, USA. Electronic address: Laura.Couzens@fda.hhs.gov.
² Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Avenue, Building 72, HFM445, Silver Spring, MD 20993, USA. Electronic address: Jin.Gao@fda.hhs.gov.
³ Department of Viroscience, Erasmus Medical Center, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands. Electronic address: k.westgeest@erasmusmc.nl.
⁴ Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Avenue, Building 72, HFM445, Silver Spring, MD 20993, USA. Electronic address: sandbult@iastate.edu.
⁵ Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Avenue, Building 72, HFM445, Silver Spring, MD 20993, USA. Electronic address: Vladimir.Lugovtsev@fda.hhs.gov.
⁶ Department of Viroscience, Erasmus Medical Center, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands. Electronic address: r.fouchier@erasmusmc.nl.
⁷ Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Avenue, Building 72, HFM445, Silver Spring, MD 20993, USA. Electronic address: Maryna.Eichelberger@fda.hhs.gov.

PMID: 25233882
DOI: 10.1016/j.jviromet.2014.09.003

Abstract

Antibodies to neuraminidase (NA), the second most abundant surface protein on influenza virus, contribute toward protection against influenza. The traditional thiobarbituric acid (TBA) method to quantify NA inhibiting antibodies is cumbersome and not suitable for routine serology. An enzyme-linked lectin assay (ELLA) described by Lambre et al. (1990) is a practical alternative method for measuring NA inhibition (NI) titers. This report describes optimization of the ELLA for measuring NI titers in human sera against influenza A viruses, using H6N1 and H6N2 viruses as antigens. The optimized ELLA is subtype-specific and reproducible. While the titers measured by ELLA are somewhat greater than those measured by a miniaturized TBA method, seroconversion rates are the same, suggesting similarity in assay sensitivity under these optimized conditions. The ELLA described in this report provides a practical format for routine evaluation of human antibody responses to NA.

Keywords: Antibody; Enzyme-linked lectin assay; Influenza; Neuraminidase; Serology.

Published by Elsevier B.V.

Publication types

Comparative Study
Validation Study

MeSH terms

Animals
Antibodies, Viral / blood*
Antibody Formation
Cross Reactions
Ferrets
Humans
Immunoenzyme Techniques
Influenza A virus / enzymology
Influenza A virus / immunology*
Influenza Vaccines
Influenza, Human / immunology*
Influenza, Human / virology
Lectins / immunology*
Neuraminidase / immunology*
Sigmodontinae
Thiobarbiturates
Viral Proteins / antagonists & inhibitors
Viral Proteins / immunology

Substances

Antibodies, Viral
Influenza Vaccines
Lectins
Thiobarbiturates
Viral Proteins
Neuraminidase
thiobarbituric acid