Assays for influenza virus hemagglutinins

A technology for influenza virus and samples, which is applied in the experimental field of influenza virus hemagglutinin, such as analyzing vaccines, can solve the problem of uncertain detection of inactivated vaccine antigens

Inactive Publication Date: 2014-04-30
NOVARTIS AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This treatment irreversibly cross-links HA, so it is uncertain whether the described 2D-HPLC method is suitable for detection of bulk inactivated vaccine antigens

Method used

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  • Assays for influenza virus hemagglutinins
  • Assays for influenza virus hemagglutinins
  • Assays for influenza virus hemagglutinins

Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0106] RP-HPLC detection of influenza HA as a method for the quantification of batches ("single batches") of monovalent influenza virus antigens. It was found that when the single batch had high specific purity and stable HA, RP-HPLC was able to quantify HA well, and the quantitative results closely matched the standard SRID results. But in cases where the vaccine contained significant amounts of denatured HA, the RP-HPLC method no longer matched the SRID assay.

[0107] For example, the table below shows the results of four A / H3N2 single batches. Total protein concentration ([mu]g / ml) was assessed by BCA, followed by HA concentration ([mu]g / ml) by SRID (standard protocol) and RP-HPLC. RP-HPLC on 2.1mm x 100mm Poros TM Performed on a R1 / 10 column operating at 60°C and a flow rate of 0.8ml / min. The mobile phases were: (A) 0.1% TFA in water, 5% acetonitrile; and (B) 0.1% TFA in 100% acetonitrile (solvent B), the A / B mixture was changed from 20% in 6.5 minutes / 80% becomes 0...

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Abstract

A combination of ultrafiltration and HPLC is used to analyze influenza virus. This combination is able to quantify hemagglutinin (HA) and correlates well with single radial immunodiffusion (SRID) results, but can be performed without the delay of waiting for immunochemical SRID reagents.

Description

[0001] This patent application claims priority to US Provisional Patent Application No. 61 / 217,405, filed May 29, 2009, the entire contents of which are incorporated herein by reference. technical field [0002] The present invention relates to the field of testing influenza virus hemagglutinins such as assay vaccines. Background technique [0003] The standard test for hemagglutinin (HA) content in inactivated influenza vaccines is based on single radial immunodiffusion ("SRID") [1, 2], which was recommended by WHO in 1978 to replace the hemagglutination-based test. Although the SRID assay is well established, it is slow, has poor dynamic range, has significant variability, and takes a long time to prepare the desired specific anti-HA serum and then calibrate it. A non-SRID assay for the quantification of influenza HA has therefore been sought. [0004] One method that has been used is reverse phase high performance liquid chromatography (RP-HPLC). For example, reference ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/16A61K39/145C07K1/20C07K1/34C07K14/11G01N33/68
CPCG01N33/68A61K39/145C12N2760/16134C12N7/00C12N2760/16151G01N2333/11A61K39/12A61P31/16A61P31/18A61P37/04C07K1/20C07K1/34C07K14/11
Inventor P·道米策Y·闻G·帕尔玛P·赖尼拉P·吴
Owner NOVARTIS AG
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