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Clinical and Vaccine Immunology, March 2006, p. 356-360, Vol. 13, No. 3
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.3.356-360.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Critical Differences between Pneumococcal Polysaccharide Enzyme-Linked Immunosorbent Assays with and without 22F Inhibition at Low Antibody Concentrations in Pediatric Sera

Isabelle Henckaerts,1* David Goldblatt,2 Lindsey Ashton,2 and Jan Poolman1

GlaxoSmithKline Biologicals, Rixensart, Belgium,1 Institute of Child Health, London, United Kingdom2

Received 29 September 2005/ Returned for modification 9 November 2005/ Accepted 18 January 2006

A comparative study was conducted between two laboratories in order to evaluate the differences between two enzyme-linked immunosorbent assay (ELISA) techniques for the detection of pneumococcal anti-capsular polysaccharide antibodies. One laboratory used an assay including heterologous 22F polysaccharide inhibition, and the other laboratory employed a non-22F reference assay. After conjugate immunization, 30 pediatric post-primary immunization sera with antipolysaccharide concentrations ranging from <0.05 to 15 µg/ml were analyzed. Aggregate reverse cumulative distribution curves combining concentrations of antibodies against serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F revealed similar results for both methods at antibody levels of >1 µg/ml. However, at antibody levels of <1 µg/ml, the distribution curve measured with the 22F inhibition ELISA shifted toward lower levels. This observation suggests that the 22F inhibition assay is more specific at low antibody concentrations, which was confirmed by heterologous polysaccharide inhibition experiments. Translation of low antibody levels suggested that the proposed threshold concentration of 0.35 µg/ml determined with the non-22F ELISA corresponded to a concentration of 0.20 µg/ml with the 22F inhibition ELISA. Pneumococcal antipolysaccharide ELISA including 22F inhibition can be recommended as a reference method.

* Corresponding author. Mailing address: Research & Development, GlaxoSmithKline Biologicals, Rue de l'Institut 89, 1330 Rixensart, Belgium. Phone: 32-(0)2-656 7648. Fax: 32-(0)2-656 9144. E-mail: isabelle.henckaerts{at}gskbio.com.

Clinical and Vaccine Immunology, March 2006, p. 356-360, Vol. 13, No. 3
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.3.356-360.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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