Validation of Automated Dilution and Transfer Methods for Rapid Assessment of Functional Immune Responses to Meningococcal Vaccines

Angela M. Payne, Xu Liu, Michael J. Caulfield, Jon H. Heinrichs

Vaccines Basic Research, Merck Sharp and Dohme Corp., West Point, USA.
Vaccines Basic Research, Merck Sharp and Dohme Corp., West Point, USA..

DOI: 10.4236/wjv.2013.33014                

Abstract

A high-throughput Serum Bactericidal Assay (SBA) was developed to monitor the functional antibody responses to capsular polysaccharide antigens from multiple serotypes of Neisseria meningitidis. This assay measures the ability of an antibody, aided by complement, to mediate killing of bacteria. Functional assays of this type are increasingly used in the quality control arena as potency release tests. Consequently, there has been an enhanced requirement for reproducibility in the performance of this type of assay. Assay validation is, therefore, important to facilitate understanding of the significance of values obtained, and to enable appropriate and informed use of the assay. This study involved the evaluation of the high-throughput serum bactericidal assay including the effects of different dilution and transfer techniques on assay performance. The results presented here demonstrate the repeatability, and the precision and ruggedness of the assay.

Keywords

Serum Bactericidal Assay; SBA; Neisseria meningitides; Automation; Repeatability

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	E. C. Gotschlich, I. Goldschneider and M. S. Artenstein, “Human Immunity to the Meningococcus V. The Effect of Immunization with Meningococcal Group C Polysaccharide on the Carrier State,” The Journal of Experimental Medicine, Vol. 129, No. 6, 1969, pp. 1385-1395. doi:10.1084/jem.129.6.1385
[2]	H. Keyserling, T. Papa, K. Koranyi, R. Ryall, E. Bassily, M. J. Bybel, K. Sullivan, G. Gilmet and A. Reinhardt, “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 Pediatrics and Adolescent Medicine, Vol. 159, No. 10, 2005, pp. 907-913. doi:10.1001/archpedi.159.10.907
[3]	S. E. Maslanka, L. L. Gheesling, D. E. Libutti, K. B. Donaldson, H. S. Harakeh, J. K. Dykes, F. F. Arhin, S. J. Devi, C. E. Frasch, J. C. Huang, P. Kriz-Kuzemenska, R. D. Lemmon, M. Lorange, C. C. Peeters, S. Quataert, J. Y. Tai and G. M. Carlone, “Standardization and a Multi-laboratory Comparison of Neisseria meningitidis Serogroup A and C Serum Bactericidal Assays. The Multi-laboratory Study Group,” Clinical and Diagnostic Laboratory Immunology, Vol. 4, No. 2, 1997, pp. 156-167.
[4]	X. Liu, S. Wang, L. Sendi and M. J. Caulfield, “High-Throughput Imaging of Bacterial Colonies Grown on Filter Plates with Application to Serum Bactericidal Assays,” Journal of Immunological Methods, Vol. 292, No. 1-2, 2004, pp. 187-193. doi:10.1016/j.jim.2004.06.021
[5]	L. E. Little, “Validation of Immunological and Biological Methods,” Biopharm-the Technology & Business of Bi pharmaceuticals, Vol. 8, No. 9, 1995, pp. 36-42.