CVI Accepts, published online ahead of print on 21 January 2009

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Clin. Vaccine Immunol. doi:10.1128/CVI.00415-08
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Optimization and Validation of a Multiplex, Electrochemiluminescence-based Detection Assay for the Quantitation of IgG Serotype-Specific Anti-Pneumococcal Antibodies in Human Serum

Rocio D. Marchese^*, Derek Puchalski, Pamela Miller, Joseph Antonello, Olivia Hammond, Tina Green, Leonard J. Rubinstein, Michael J. Caulfield, and Daniel Sikkema

Wayne Clinical Support, Merck Research Laboratories, Wayne, Pennsylvania; Non-Clinical Statistics, Merck Research Laboratories, West Point, Pennsylvania; ID/Vaccines, Merck Research Laboratories, Upper Gwynedd, Pennsylvania

^* To whom correspondence should be addressed. Email: rocio_marchese{at}merck.com.

PNEUMOVAX® 23 consists of a mixture of highly purified capsular polysaccharides (Ps) from 23 of the most prevalent serotypes of Streptococcus pneumoniae (Pn). Testing of vaccine immunogenicity has been historically performed on the ELISA platform, validated to measure IgG antibodies to all 23 serotypes included in PNEUMOVAX® 23. In order to significantly improve the throughput of this testing, we have developed and validated a direct binding electrochemiluminescence (ECL)-based multiplex assay that can measure antibody response in human serum to eight serotypes within a single microtiter well. The Pn ECL assay is based on the Meso Scale Discovery (MSD) technology which utilizes a SULFO-TAG^TM labeled anti-human IgG antibody that emits light upon electrochemical stimulation. The Pn ECL assay exhibits a wide dynamic range and provides for the ability to read concentrations down to the minimum reported concentration in the Merck ELISA of 0.1 µg/ml. Cross-reactivity assessment using type specific monoclonal antibodies showed no "cross talk" between antigen spots within a well. Using the WHO Pneumococcal sample reference panel, the results obtained in the Pn ECL assay were compared to the results obtained in the international Pn ELISA. The results for the Pn ECL satisfied the WHO recommended acceptance criterion for concordance for all seven serotypes with published Pn ELISA values and the overall correlation (r value) across the 7 serotypes was 0.994. The MSD methodology has a high potential to be extremely useful for simultaneously quantitating IgG responses to several Pneumococcal serotypes, while conserving serum volume and laboratory testing time.