A convenient enzyme-linked immunosorbent assay for rapid screening of anti-adeno-associated virus neutralizing antibodies

This version was published on 1 November 2009

Ann Clin Biochem 2009;46:508-510
© 2009 Association for Clinical Biochemistry


This Article
Right arrow
Full Text
Right arrow

Full Text (PDF)

Right arrow
All Versions of this Article:



most recent

Right arrow
Alert me when this article is cited
Right arrow
Alert me if a correction is posted
Right arrow
Email this article to a friend
Right arrow

Similar articles in this journal

Right arrow
Similar articles in PubMed
Right arrow
Alert me to new issues of the journal
Right arrow
Download to citation manager
Right arrow
Google Scholar
Right arrow
Articles by Ito, T.
Right arrow
Articles by Muramatsu, S.-i.
Right arrow
PubMed Citation
Social Bookmarking

What’s this?

Short Reports

Tetsuo Ito1,
Shigekazu Yamamoto1,
Tsukasa Hayashi1,
Mika Kodera1,3,
Hiroaki Mizukami2,
Keiya Ozawa2 and
Shin-ichi Muramatsu3

1 KAINOS Laboratories Inc., Tokyo;
2 Division of Genetic Therapeutics;
3 Division of Neurology, Jichi Medical University, Tochigi, Japan

Corresponding author: Shin-ichi Muramatsu, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan. Email: muramats{at}jichi.ac.jp

Background: Recombinant adeno-associated virus vectors based on serotype2 (AAV-2) have become leading vehicles for gene therapy. Mosthumans in the general population have anti-AAV-2 antibodiesas a result of naturally acquired infections. Pre-existing immunityto AAV-2 might affect the functional and safety consequencesof AAV-2 vector-mediated gene transfer in clinical applications.

Methods: An enzyme-linked immunosorbent assay (ELISA) method was developedusing microwell plates coated with intact particles of recombinantAAV-2 vectors, and horseradish peroxidase-conjugated anti-humanimmunoglobulin G (HRP-IgG). Neutralizing antibody titres wereanalysed by assessing the ability of serum antibody to inhibittransduction into HEK293 cells of AAV vectors that express β-galactosidase.

Results: Anti-AAV-2 antibodies were detected by ELISA in two of 20 healthysubjects. The positivity criterion (optical density >0.5)in ELISA corresponded to the cut-off value (320-fold dilutionof serum) in the AAV-2 neutralization assay. Influences of interferingsubstances were not observed.

Conclusion: This ELISA method may be useful for rapid screening of anti-AAV-2neutralizing antibodies in candidates for gene therapy.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What’s this?