Multiplexed VaxArray immunoassay for rapid antigen quantification in measles and rubella vaccine manufacturing

Jacob H Gillis,Keely N Thomas,Senthilkumar Manoharan,Mallikarjuna Panchakshari,Amber W Taylor,David F Miller,Rose T Byrne-Nash,Christine Riley,Kathy L Rowlen,Erica Dawson

doi:10.1016/j.jvacx.2021.100113

Jacob H Gillis, Keely N Thomas + Show 8 more

Open Access

https://doi.org/10.1016/j.jvacx.2021.100113

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Journal: Vaccine: X	Publication Date: Sep 20, 2021
Citations: 5	License type: cc-by

Affiliation: Biological E (India), InDevR (United States)

Abstract

Measles-containing vaccines (MCV), specifically vaccines against measles and rubella (MR), are extremely effective and critical for the eradication of measles and rubella diseases. In developed countries, vaccination rates are high and vaccines are readily available, but continued high prevalence of both diseases in developing countries and surges in measles deaths in recent years have highlighted the need to expand vaccination efforts. To meet demand for additional vaccines at a globally affordable price, it is highly desirable to streamline vaccine production thereby reducing cost and speeding up time to delivery. MR vaccine characterization currently relies on the 50% cell culture infectious dose (CCID50) assay, an endpoint assay with low reproducibility that requires 10–14 days to complete. For streamlining bioprocess analysis and improving measurement precision relative to CCID50, we developed the VaxArray Measles and Rubella assay kit, which is based on a multiplexed microarray immunoassay with a 5-hour time to result. Here we demonstrate vaccine-relevant sensitivity ranging from 345 to 800 IFU/mL up to 100,000 IFU/mL (infectious units per mL) and specificity that allows simultaneous analysis in bivalent vaccine samples. The assay is sensitive to antigen stability and has minimal interference from common vaccine additives. The assay exhibits high reproducibility and repeatability, with 15% CV, much lower than the typical 0.3 log10 error (∼65%) observed for the CCID50 assay. The intact protein concentration measured by VaxArray is reasonably correlated to, but not equivalent to, CCID50 infectivity measurements for harvest samples. However, the measured protein concentration exhibits equivalency to CCID50 for more purified samples, including concentrated virus pools and monovalent bulks, making the assay a useful new tool for same-day analysis of vaccine samples for bioprocess development, optimization, and monitoring.

Highlights

Measles and rubella are highly contagious viruses that cause significant morbidity and mortality
Acute measles infection is associated with fever, cough, and rash that can persist for a week [2]
A panel of 21 monoclonal antibodies, raised against measles (n = 9) or rubella (n = 12) were each printed in triplicate on the microarray Antibodies were evaluated for reactivity, specificity, limit of detection, linear dynamic range, and stability indicating capabilities, with a single antibody for each virus was down-selected for inclusion in the final assay

Summary

Introduction

Measles and rubella are highly contagious viruses that cause significant morbidity and mortality. Measles virus is one of the most infectious human-borne viruses and, prior to widespread vaccination, infected > 90% of children before 15 years of age [1]. Acute measles infection is associated with fever, cough, and rash that can persist for a week [2]. Up to 6% of acute measles cases are fatal and the World Health Organization (WHO) estimates 140,000 children, most in the developing world, died from acute measles infection in 2018 [3]. Complication of measles infection, can occur in patients up to 10 years after initial acute infection. While rare (0.007–0.011% of infections [4]), SSPE is always fatal, usually within 1 to 3 years

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