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Journal of Clinical Microbiology, November 2003, p. 5046-5052, Vol. 41, No. 11
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.11.5046-5052.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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

Mieke C. Sprangers, Wandena Lakhai, Wouter Koudstaal, Marielle Verhoeven, Björn F. Koel, Ronald Vogels, Jaap Goudsmit, Menzo J. E. Havenga, and Stefan Kostense*

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

Received 7 November 2002/ Returned for modification 29 March 2003/ Accepted 1 September 2003

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.

* Corresponding author. Mailing address: Crucell Holland BV, P.O. Box 2048, 2301 CA Leiden, The Netherlands. Phone: 31 (0) 71 5248834. Fax: 31 (0) 71 5248702. E-mail: S.Kostense{at}crucell.com.

Journal of Clinical Microbiology, November 2003, p. 5046-5052, Vol. 41, No. 11
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.11.5046-5052.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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