Quantifying Adenovirus-Neutralizing Antibodies by Luciferase Transgene Detection: Addressing Preexisting Immunity to Vaccine and Gene Therapy Vectors

Author:

Sprangers Mieke C.1,Lakhai Wandena1,Koudstaal Wouter1,Verhoeven Marielle1,Koel Björn F.1,Vogels Ronald1,Goudsmit Jaap1,Havenga Menzo J. E.1,Kostense Stefan1

Affiliation:

1. Vaccine R&D Division, Crucell Holland BV, 2301 CA Leiden, The Netherlands

Abstract

ABSTRACT The presence of various levels of anti-adenovirus serotype 5 (Ad5)-neutralizing antibodies in humans is thought to contribute to the inconsistent clinical results obtained so far in diverse gene transfer and vaccination studies and might preclude universal dosing with recombinant Ad5. Prescreening of individuals eligible for Ad5 or alternative serotype treatment and subsequently tailoring the vector dose might aid in ensuring the consistency of clinical parameters. For this purpose, a qualified Ad neutralization assay is required. Here we have tested the different protocols used to date to determine anti-Ad neutralizing activity. Based on simplicity, speed, high throughput, sensitivity, and robustness, we propose a qualified assay in which Ad neutralization is monitored by luciferase reporter gene expression.

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

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