Development of a Multi-Plex Electrochemiluminescent Assay for the Detection of Serum Antibody Responses to Meningococcal Conjugate Vaccines

DOI: 10.4236/wjv.2012.21004   PDF   HTML     4,105 Downloads   8,121 Views  


Neisseria meningitidis is a gram negative diplococcal bacterium. Worldwide, N. meningitidis is the leading cause of bacterial meningitis and sepsis, with five serogroups (A, B, C, Y, and W-135) responsible for the majority of the disease. Multivalent (A, C, Y, and W-135) polysaccharide and conjugate vaccines have been licensed in the United States and elsewhere and are widely available. We have developed a multi-plexed electrochemiluminescent assay to quantitate serum antibody responses to meningococcal polysaccharides A, C, Y, and W-135 to allow for rapid evaluation of li- censed and investigational vaccines. A 96-well plate containing a carbon electrode arrayed with polysaccharides A, C, Y, and W-135 on separate spots within each well has been developed for simultaneous detection of polysaccharidespecific antibodies in serum samples from vaccinated individuals. The assay conditions were optimized using the anti-meningococcal serogroup A/C reference serum pool, CDC 1992 (NIBSC 99/706), through evaluation of plate types, coating polysaccharide concentrations, and blocking and serum diluent buffers. Comparison of single and multiplex assays demonstrated the sensitivity, specificity, and speed of the multi-plex format for the quantification of serum antibody responses to N. meningitidis polysaccharides A, C, Y and W-135.

Share and Cite:

L. Indrawati, J. Heinrichs, E. Wen and J. Skinner, "Development of a Multi-Plex Electrochemiluminescent Assay for the Detection of Serum Antibody Responses to Meningococcal Conjugate Vaccines," World Journal of Vaccines, Vol. 2 No. 1, 2012, pp. 27-35. doi: 10.4236/wjv.2012.21004.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] F. M. LaForce, et al., “The Meningitis Vaccine Project,” Vaccine, Vol. 25, Supplement 1, 2007, pp. 97-100. doi:10.1016/j.vaccine.2007.04.049
[2] M. E. Pichichero, “Meningococcal Conjugate Vaccines,” Expert Opinion on Biological Therapy, Vol. 5, No. 11, 2005, pp. 1475-89. doi:10.1517/14712598.5.11.1475
[3] M. Broker and S. Fantoni, “Meningococcal Disease: A Review on available Vaccines and Vaccines in Development,” Minerva Medica, Vol. 98, No. 5, 2007, pp. 575-89.
[4] J. D. Campbell, et al., “Safety, Reactogenicity, and Immunogenicity of a Tetravalent Meningococcal Polysaccharide-Diphtheria Toxoid Conjugate Vaccine Given to Healthy Adults,” The Journal of Infectious Diseases, Vol. 186, No. 12, 2002, pp. 1848-1851. doi:10.1086/345763
[5] B. Cooper, L. DeTora and J. Stoddard, “Menveo?: A Novel Quadrivalent Meningococcal CRM197 Conjugate Vaccine against Serogroups A, C, W-135 and Y,” Expert Reviews of Vaccines, Vol. 10, No. 1, 2011, pp. 21-33. doi:10.1586/erv.10.147
[6] M. Rennels, et al., “Dosage Escalation, Safety and Immunogenicity Study of Four Dosages of a Tetravalent Meninogococcal Polysaccharide Diphtheria Toxoid Conjugate Vaccine in Infants,” Pediatric Infectious Disease Journal, Vol. 23, No. 5, 2004, pp. 429-35. doi:10.1097/01.inf.0000126297.28952.f8
[7] M. D. Snape, et al., “Immunogenicity of a Tetravalent Meningococcal Glycoconjugate Vaccine in Infants: a Randomized Controlled Trial,” The Journal of the American Medical Association, Vol. 299, No. 2, 2008, pp. 173-84. doi:10.1001/jama.2007.29-c
[8] G. F. Blackburn, et al., “Electrochemiluminescence Detection for Development of Immunoassays and DNA Probe Assays for Clinical Diagnostics,” Clinical Chemistry, Vol. 37, No. 9, 1991, pp. 1534-1539.
[9] V. Guglielmo-Viret and P. Thullier, “Comparison of an Electrochemiluminescence Assay in Plate Format over a Colorimetric ELISA, for the Detection of Ricin B Chain (RCA-B),” Journal of Immunological Methods, Vol. 328, No. 1-2, 2007, pp. 70-78. doi:10.1016/j.jim.2007.08.003
[10] R. M. de Voer, et al., “Development of a FluorescentBead-Based Multiplex Immunoassay to Determine Immunoglobulin G Subclass Responses to Neisseria meningitidis Serogroup A and C Polysaccharides,” Clinical and Vaccine Immunology, Vol. 15, No. 8, 2008, pp. 1188-93. doi:10.1128/CVI.00478-07
[11] T. B. Martins, et al., “Development of a Multiplexed Fluorescent Immunoassay for the Quantitation of Antibody Responses to Four Neisseria meningitidis Serogroups,” Journal of Immunological Methods, Vol. 342, No. 1-2, 2009, pp. 98-105. doi:10.1016/j.jim.2008.12.003
[12] R. D. Marchese, et al., “Optimization and Validation of a Multiplex, Electrochemiluminescence-Based Detection Assay for the Quantitation of IgG Serotype-Specific Antipneumococcal Antibodies in Human Serum,” Clinical and Vaccine Immunology, Vol. 16, No. 3, 2009, pp. 387-396. doi:10.1128/CVI.00415-08
[13] C. E. Frasch, “Production and Control of Neisseria meningitidis Vaccines,” Advances in Biotechnological Processes, Vol. 13, 1990, pp. 123-145.
[14] A. L. Lee, M. S. Rienstra, W. E. Manger and R. D. Sitrin, “Process for converting Lipid-Containing Bacterial Capsular Polysaccharide into Lipid-Free Polysaccharide,” Merck & Co., Rahway, 1994.
[15] P. K. Holder, et al., “Assignment of Neisseria meningitisdis Serogroup A and C Class-Specific Anticapsular Antibody Concentrations to the New Standard Reference Serum CDC1992,” Clinical and Diagnostic Laboratory Immunology, Vol. 2, No. 2, 1995, pp. 132-137.
[16] D. C. Phipps, et al., “Standardization of ELISA for the Quantitation of Antibodies to S. pneumoniae Capsular Polysaccharides (PnPs),” Pediatric Research, Vol. 27, 1990, p. 179A.
[17] S. A. Quataert, et al., “Assignment of Weight-Based Antibody Units to a Human Antipneumococcal Standard Reference Serum, Lot 89-S,” Clinical and Diagnostic Laboratory Immunology, Vol. 2 No. 5, 1995, pp. 590-597.
[18] G. M. Carlone, et al., “Multicenter Comparison of Levels of Antibody to the Neisseria meningitidis Group A Capsular Polysaccharide Measured by Using an Enzyme-Linked Immunosorbent Assay,” Journal of Clinical Microbiology, Vol. 30, No. 1, 1992, pp. 154-159.
[19] A. J. Pollard and M. C. J. Maiden, “Meningococcal Disease: Methods and Protocols,” In: J. M. Walker, Ed., Methods in Molecular Medicine, Vol. 67, Humana Press, Totowa, 2001.
[20] Advisory Committee on Immunization Practices (ACIP), “Licensure of a Meningococcal Conjugate Vaccine (Menveo) and Guidance for Use,” Morbidity and Mortality Weekly Report, Vol. 59, No. 9, 2010, p. 273.
[21] D. R. Deaver, “A New Non-Isotopic Detection System for Immunoassays,” Nature, Vol. 377, No. 6551, 1995, pp. 758-760. doi:10.1038/377758a0
[22] H. Joseph, et al., “Assignment of Neisseria meningitidis Serogroups A, C, W135, and Y Anticapsular Total Immunoglobulin G (IgG), IgG1, and IgG2 Concentrations to Reference Sera,” Clinical and Diagnostic Laboratory Immunology, Vol. 11, No. 1, 2004, pp. 1-5.
[23] R. D. Marchese, et al., “Enzyme-Linked Immunosorbent Assay for Measuring Antibodies to Pneumococcal Polysaccharides for the PNEUMOVAX 23 Vaccine: Assay Operating Characteristics and Correlation to the WHO International Assay,” Clinical and Vaccine Immunology, Vol. 13, No. 8, 2006, pp. 905-912. doi:10.1128/CVI.00014-06
[24] D. R. Bundle, I. C. Smith and H. J. Jennings, “Determination of the Structure and Conformation of Bacterial Polysaccharides by Carbon 13 Nuclear Magnetic Resonance. Studies on the Group-Specific Antigens of Neisseria meningitidis Serogroups A and X,” The Journal of Biological Chemistry, Vol. 249, No. 7, 1974, pp. 2275-2281.
[25] C. W. Chen and J. S. Cohen, “Salt- and Sequence-Dependence of the Secondary Structure of DNA in Solution by 31P-NMR Spectroscopy,” Biopolymers, Vol. 22, No. 3, 1983, pp. 879-893. doi:10.1002/bip.360220310
[26] M. Vorlickova, J. Kypr and V. Sklenar, “Salt-Induced Conformational Transition of Poly[d(A-T)]-Poly[d(A-T)],” Journal of Molecular Biology, Vol. 166, No. 1, 1983, pp. 85-92. doi:10.1016/S0022-2836(83)80052-7
[27] A. Kalra, N. Tugcu, S. M. Cramer and S. Garde, “Salting-In and Salting-Out of Hydrophobic Solutes in Aqueous Salt Solutions,” The Journal of Physical Chemistry B, Vol. 105, No. 27, 2001, pp. 6380-6386. doi:10.1021/jp010568+
[28] M. Foschiatti, et al., “Conformational Studies of the Capsular Polysaccharide Produced by Neisseria meningitidis group A,” Carbohydrate Research, Vol. 344, No. 7, 2009, pp. 940-943. doi:10.1016/j.carres.2009.02.027
[29] A. K. Bhattacharjee, et al., “Structural Determination of the Polysaccharide Antigens of Neisseria meningitidis Serogroups Y, W-135, and BO1,” Canadian Journal of Biochemistry, Vol. 54, No. 1, 1976, pp. 1-8. doi:10.1139/o76-001
[30] C. M. John, M. Liu and G. A. Jarvis, “Profiles of Structural Heterogeneity in Native Lipooligosaccharides of Neisseria and Cytokine Induction,” Journal of Lipid Research, Vol. 50, No. 3, 2009, pp. 424-38. doi:10.1194/jlr.M800184-JLR200
[31] H. Keyserling, et al., “Safety, Immunogenicity, and Immune Memory of a Novel Meningococcal (Groups A, C, Y, and W-135) Polysaccharide Diphtheria Toxoid Conjugate vaccine (MCV-4) in Healthy Adolescents,” Archives of Pediatrics & Adolescent Medicine, Vol. 159, No. 10, 2005, pp. 907-913. doi:10.1001/archpedi.159.10.907

comments powered by Disqus

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.