Development and Standardization of Elisa for Identification and Quantitation of Neisseria Meningitidis Antigens for Vaccine Development

Abstract

The purpose of this study was to develop and standardize immunoassay for newlineidentification and quantification of meningococcal PS at various stages of vaccine development. The study included the development of in-house primary antibodies against Neisseria meningitidis serogroup A (MenA), serogroup W (MenW) and newlineserogroup X (MenX). These in-house antibodies were then utilized for the development and standardization of competitive/inhibition ELISA, an immunoassay to identify and newlinequantify capsular PS of MenA, W and X serogroups. This PhD work further included newlineoptimization of upstream and downstream processes to improve the yield of newlinemeningococcal PS. The study also involved the evaluation of impact of pH and newlinetemperature alterations on the size and antigenicity of meningococcal serogroup A and X PSs and conjugates. Elevated temperature and the change in pH may result in the physical instability leading to the size degradation of the PS. Moreover, high temperature may also result in protein aggregation and altered tertiary structure of the protein in the conjugate. In our study we have used inhibition ELISA to assess the impact of temperature and pH alterations on the antigenicity of N. meningitidis serogroup A and X, PS and conjugates and its correlation with the size distribution analysis using high pressure size exclusion chromatography. This study is an attempt to reduce the production cost of multivalent vaccines and the final cost of the vaccine in newlinethe market in the developing countries like India. There is a need to improve the newlinetechnologies for the production of affordable and efficient vaccines against newlinemeningococcal meningitis in the world.PS is an important antigen for the vaccine and therefore it is important to study its production, purification, conjugation and in vaccine formulation. These steps are extremely complex and therefore competitive ELISA being developed helps to identify and estimate the PS content for the vaccine formulation. newline newline