A neuraminidase potency assay for quantitative assessment of neuraminidase in influenza vaccines

Byrne-Nash, Rose T.; Gillis, Jacob H.; Miller, David F.; Bueter, Katie M.; Kuck, Laura R.; Rowlen, Kathy L.

doi:10.1038/s41541-019-0099-3

Cited by 7 publications

(8 citation statements)

References 41 publications

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“…The VaxArray Measles and Rubella (MR) assay is similar to previously-described VaxArray Influenza assays [21] , [22] , [23] , [24] , with the slide layout, microarray layout, and detection principle depicted in Fig. 1 a, b, and c , respectively.…”

Section: Methodsmentioning

confidence: 99%

“…This work describes the development of an alternative assay for tracking antigen content, stability, and identity throughout the MR vaccine manufacturing process. We have adapted the VaxArray technology, described previously for influenza vaccines [21] , [22] , [23] , [24] , for the evaluation of MR vaccine antigens. VaxArray assays are simple multiplexed immunoassays that use monoclonal antibodies in a glass microarray format.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2021

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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 (CCID 50 ) assay, an endpoint assay with low reproducibility that requires 10–14 days to complete. For streamlining bioprocess analysis and improving measurement precision relative to CCID 50 , 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 log 10 error (∼65%) observed for the CCID 50 assay. The intact protein concentration measured by VaxArray is reasonably correlated to, but not equivalent to, CCID 50 infectivity measurements for harvest samples. However, the measured protein concentration exhibits equivalency to CCID 50 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.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2021

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“…Given the rapid timelines for vaccine development already underway, a multiplexed assay that can measure vaccine-induced antibody response to a variety of related antigens simultaneously is highly desirable for both time and cost savings. The VaxArray platform (InDevR, Inc., Boulder, CO) is a microscale, multiplexed, microarray-based immunoassay platform that has been well-validated for use in influenza vaccine antigen characterization ( Kuck et al, 2018 ; Byrne-Nash et al, 2019 ), and has been adapted for serological analysis of coronaviruses with the recent availability of the Coronavirus (CoV) SeroAssay. Specifically, nine unique coronavirus spike protein antigens are printed in replicate in a microarray format, providing the ability to perform simultaneous analysis of antibody responses to all 9 antigens in a single, 2 -h assay.…”

Section: Introductionmentioning

confidence: 99%

Multiplexed, microscale, microarray-based serological assay for antibodies against all human-relevant coronaviruses

Dawson

Kuck

Blair

et al. 2021

Journal of Virological Methods

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Rapid, sensitive, and precise multiplexed assays for serological analysis during candidate COVID-19 vaccine development would streamline clinical trials. The VaxArray Coronavirus (CoV) SeroAssay quantifies IgG antibody binding to 9 pandemic, potentially pandemic, and endemic human CoV spike antigens in 2 hours with automated results analysis. IgG antibodies in serum bind to the CoV spike protein capture antigens printed in a microarray format and are labeled with a fluorescent anti-species IgG secondary label. The assay demonstrated excellent lower limits of quantification ranging from 0.3 – 2.0 ng/mL and linear dynamic ranges of 76 to 911-fold. Average precision of 11% CV and accuracy (% recovery) of 92.5% over all capture antigens were achieved over 216 replicates representing 3 days and 3 microarray lots. Clinical performance on 263 human serum samples (132 SARS-CoV-2 negatives and 131 positives based on donor-matched RT-PCR and/or date of collection) produced 98.5% PPA and 100% NPA.

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“…MR vaccines rely on cell-culture based assays, such as 50% Cell Culture Infectious Dose (CCID50), for characterization during manufacturing, including identity, potency, and stability testing. Infectivity measurements are widely adopted and highly predictive of vaccine efficacy [17][18][19][20] but present certain limitations, as the time to result is 10-14 days and execution requires trained personnel. In addition, CCID50 assays rely on subjective, discontinuous endpoint measurements and are notoriously error prone with observed variability of ± 0.3 log10 infectious units per mL (50-100% variability) [21][22][23], resulting in costly hold times and lot rejections that can increase manufacturing costs and delay delivery of critical vaccine doses.…”

Section: Introductionmentioning

confidence: 99%

Multiplexed VaxArray Immunoassay for Rapid Antigen Quantification in Measles and Rubella Vaccine Manufacturing

Gillis¹,

Thomas

Manoharan

et al. 2021

Preprint

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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 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.

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A neuraminidase potency assay for quantitative assessment of neuraminidase in influenza vaccines

Cited by 7 publications

References 41 publications

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

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

Multiplexed, microscale, microarray-based serological assay for antibodies against all human-relevant coronaviruses

Multiplexed VaxArray Immunoassay for Rapid Antigen Quantification in Measles and Rubella Vaccine Manufacturing

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