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Virus-Based Expression Systems Facilitate Rapid Target In Vivo Functionality Validation and High-Throughput Screening

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Kelly A. Conway

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Anil H. Vaidya

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Daniel Rosenthal

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Mary Jo Wildey

Raritan, NJ

Lisa Minor

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Zina Itkin

Johnson & Johnson Pharmaceutical Research & Development, Raritan, NJ

Yan Kong

Johnson & Johnson Pharmaceutical Research & Development, Raritan, NJ

Jennifer Piesvaux

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Jenson Qi

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Marc Mercken

Johnson & Johnson Pharmaceutical Research & Development, Beerse, Belgium

Patricia Andrade-Gordon

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Carlos Plata-Salamán

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA

Sergey E. Ilyin

Johnson & Johnson Pharmaceutical Research & Development, Spring House, PA, silyin{at}prdus.jnj.com

Target validation is one of rate-limiting steps in the modern drug discovery. The authors developed a strategy of combining adenovirus-mediated gene transfer for efficient target functionality validation, both in vivo and in vitro, with baculovirus expression to produce sufficient quantities of protein for high-throughput screening (HTS). The incorporation of green fluorescent protein (GFP) in the adenovirus vectors accelerates recombinant adenovirus plaque purification, whereas the use of epitope and affinity tags facilitates the identification and purification of recombinant protein. In this generalized scheme, the flexible modular design of viral vectors facilitates the transition between target validation and HTS. In the example presented, functional target validation in vivo was achieved by overexpressing the target gene in cell-based models and in the mouse cortex following adenovirus-mediated gene delivery. In this context, target overexpression resulted in the accumulation of a disease-related biomarker both in vitro and in vivo. A baculovirus-based expressional system was then generated to produce enough target protein for HTS. Thus, the use of these viral expression systems represents a generalized method for rapid target functionality validation and HTS assay development, which could be applied to numerous target candidates being elucidated in gene discovery programs. (Journal of Biomolecular Screening 2003:65-71)

Key Words: Adenovirus • target validation • bioinformatics • HTS

Journal of Biomolecular Screening, Vol. 8, No. 1, 65-71 (2003)
DOI: 10.1177/1087057102239664


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