Next Article in Journal
Method for the Routine Determination of Accurate Masses by Triple Quadrupole Mass Spectrometry
Previous Article in Journal
Fast PET Scan Tumor Segmentation Using Superpixels, Principal Component Analysis and K-Means Clustering
Article Menu
Issue 1 (March) cover image

Export Article

Open AccessProtocol
Methods Protoc. 2018, 1(1), 8; https://doi.org/10.3390/mps1010008

Pseudotype Neutralization Assays: From Laboratory Bench to Data Analysis

Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Greenwich and Kent at Medway, Chatham ME4 4TB, UK
*
Author to whom correspondence should be addressed.
Received: 11 December 2017 / Revised: 15 January 2018 / Accepted: 16 January 2018 / Published: 22 January 2018
View Full-Text   |   Download PDF [9018 KB, uploaded 24 January 2018]   |  

Abstract

Pseudotype neutralization assays are powerful tools to study functional antibody responses against viruses in low biosafety laboratories. However, protocols described in the literature differ widely with respect to material, reagents, and methods used to perform these assays and to analyse the raw data generated. This could result in discrepancies between the results of different laboratories even when the same pseudotypes and the same samples are analysed. Here, we describe, in detail, an experimental protocol to perform pseudotype neutralization assays using lentiviral pseudotypes bearing influenza haemagglutinin and expressing firefly luciferase. We also present the steps necessary to analyse the data and calculate the half maximal inhibitory concentration of the sera analysed. This protocol will provide support for the validation and the standardization of the pseudotype neutralization assay for influenza virus serology. Additionally, it will provide a starting point for the development of pseudotype neutralization assays using pseudotypes bearing other viral envelope proteins. View Full-Text
Keywords: influenza; pseudotypes; serology; lentiviral vector; antibodies; neutralization influenza; pseudotypes; serology; lentiviral vector; antibodies; neutralization
Figures

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
SciFeed

Share & Cite This Article

MDPI and ACS Style

Ferrara, F.; Temperton, N. Pseudotype Neutralization Assays: From Laboratory Bench to Data Analysis. Methods Protoc. 2018, 1, 8.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Methods Protoc. EISSN 2409-9279 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top