Neuraminidase (NA) immunity leads to decreased viral shedding and reduced severity of influenza disease; however, NA content in influenza vaccines is currently not regulated, resulting in inconsistent quality and quantity of NA that can vary from manufacturer to manufacturer, from year to year, and from lot to lot. To address this problem, we have developed an assay for NA quantification that could be used by the industry to move toward developing influenza vaccines that induce a predictable immune response to NA. The VaxArray Influenza Seasonal NA Potency Assay (VXI-sNA) is a multiplexed sandwich immunoassay that relies on six subtype-specific monoclonal antibodies printed in microarray format and a suite of fluor-conjugated “label” antibodies. The performance of the assay as applied to a wide range of influenza vaccines is described herein. The assay demonstrated high NA subtype specificity and high sensitivity, with quantification limits ranging from 1 to 60 ng/mL and linear dynamic ranges of 24–500-fold. When compared to an enzymatic activity assay for samples exposed to thermal degradation conditions, the assay was able to track changes in protein stability over time and exhibited good correlation with enzyme activity. The assay also demonstrated excellent analytical precision with relative error ranging from 6 to 12% over day-to-day, user-to-user, and lot-to-lot variation. The high sensitivity and reproducibility of the assay enabled robust detection and quantification of NA in crude in-process samples and low-dose, adjuvanted vaccines with an accuracy of 100 ± 10%.

神经氨酸酶（NA）免疫力导致病毒脱落减少和流感疾病严重程度降低；但是，流感疫苗中的NA含量目前尚未受到监管，导致NA的质量和数量不一致，制造商之间，年份之间以及批次之间的NA数量和质量可能会有所不同。为了解决这个问题，我们开发了一种用于NA定量的测定方法，该方法可被业界用于开发可诱导对NA产生可预测的免疫反应的流感疫苗。VaxArray流感季节性NA效价测定（VXI-sNA）是一种多重夹心免疫测定，它依赖于以微阵列格式打印的六种亚型特异性单克隆抗体和一套与荧光缀合的“标记”抗体。本文描述了应用于多种流感疫苗的测定的性能。该测定法显示出高NA亚型特异性和高灵敏度，定量限范围为1至60 ng / mL，线性动态范围为24-500倍。与暴露于热降解条件下的样品的酶活性测定相比，该测定能够追踪蛋白质稳定性随时间的变化，并与酶活性表现出良好的相关性。该测定法还显示出出色的分析精度，相对误差在日常，用户之间和批次之间变化范围为6％至12％。该测定法的高灵敏度和可重复性可对粗制过程中的样品和小剂量佐剂疫苗中的NA进行可靠的检测和定量，准确度为100±10％。定量限范围为1至60 ng / mL，线性动态范围为24-500倍。与暴露于热降解条件下的样品的酶活性测定相比，该测定能够追踪蛋白质稳定性随时间的变化，并与酶活性表现出良好的相关性。该测定法还显示出出色的分析精度，相对误差在日常，用户之间和批次之间变化范围为6％至12％。该测定法的高灵敏度和可重复性可对粗制过程中的样品和小剂量佐剂疫苗中的NA进行可靠的检测和定量，准确度为100±10％。定量限范围为1至60 ng / mL，线性动态范围为24-500倍。与暴露于热降解条件下的样品的酶活性测定相比，该测定能够追踪蛋白质稳定性随时间的变化，并与酶活性表现出良好的相关性。该测定法还显示出出色的分析精度，相对误差在日常，用户之间和批次之间变化范围为6％至12％。该测定法的高灵敏度和可重复性可对粗制过程中的样品和小剂量佐剂疫苗中的NA进行可靠的检测和定量，准确度为100±10％。该测定法能够跟踪蛋白质稳定性随时间的变化，并显示出与酶活性的良好相关性。该测定法还显示出出色的分析精度，相对误差在日常，用户之间和批次之间变化范围为6％至12％。该测定法的高灵敏度和可重复性可对粗制过程中的样品和小剂量佐剂疫苗中的NA进行可靠的检测和定量，准确度为100±10％。该测定法能够跟踪蛋白质稳定性随时间的变化，并显示出与酶活性的良好相关性。该测定法还显示出出色的分析精度，相对误差在日常，用户之间和批次之间变化范围为6％至12％。该测定法的高灵敏度和可重复性可对粗制过程中的样品和小剂量佐剂疫苗中的NA进行可靠的检测和定量，准确度为100±10％。