English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Monitoring Influenza Virus Content in Vaccine Production : Precise Assays for the Quantitation of Hemagglutination and Neuraminidase Activity

Kalbfuss, B., Knöchlein, A., Kröber, T., & Reichl, U. (2008). Monitoring Influenza Virus Content in Vaccine Production: Precise Assays for the Quantitation of Hemagglutination and Neuraminidase Activity. Biologicals, 36(3), 145-161. doi:10.1016/j.biologicals.2007.10.002.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0013-95FA-0 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-A1FD-7
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Kalbfuss, B.1, Author              
Knöchlein, A.1, 2, Author              
Kröber, T.1, Author              
Reichl, U.1, 3, Author              
Affiliations:
1Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society, ou_1738140              
2Univ. of Applied Sciences Hamburg, Faculty of Life Sciences, persistent:22              
3Otto-von-Guericke-Universität Magdeburg, ou_1738156              

Content

show
hide
Free keywords: assay development; influenza virus; vaccine; hemagglutination activity; neuraminidase activity
 Abstract: Robust and precise quantitation of influenza virus is a premise for the efficient development of vaccine production processes. In this article, revised assays for the determination of hemagglutination (NA)and neuraminidase (NA) activity are presented. Bias of traditional discontinuous HA assays and operator dependency was overcome by introduction of a regression procedure. At little effort, a continuous assay result is obtained with repeatability as good as +29% / -22% in the best case (95% confidence intervals reported). Similarly, neuraminidase activity determined in microtiter plates resulted in repeatability better than ±20%. NA activity decreased almost linearly for pH ranging from 5.8 to 7.8 and was enhanced by the addition of Ca2+. Non-linearity of the assay (due to unspecific adsorption) was overcome by addition of BSA. Using -methylumbelliferyl- -D-Nacetylneuraminic acid as substrate, Michaelis-Menten constants of 30 and 460 μM were determined for strains A/PR/8/34 (H1N1) and A/Equi 2/NM/1/93 (H3N8), respectively. The error introduced by approximation of Michaelis-Menten kinetics (zero and first order) was minimized by limiting substrate consumption to about 10%. Linearity of both assays was verified in dilution experiments. Applicability was demonstrated in three cases: virus propagation in mammalian cell culture, ultrafiltration and precipitation of nucleic acids. 056/08/$34.00 2007 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved. [accessed 2013 November 14th]

Details

show
hide
Language(s): eng - English
 Dates: 2008
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 327756
DOI: 10.1016/j.biologicals.2007.10.002
Other: 30/08
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Biologicals
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: London : Academic Press
Pages: - Volume / Issue: 36 (3) Sequence Number: - Start / End Page: 145 - 161 Identifier: ISSN: 1045-1056
CoNE: /journals/resource/954927604698