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Utilizing the Antigen Capsid-Incorporation Strategy for the Development of Adenovirus Serotype 5-Vectored Vaccine Approaches
Published on: May 6, 2015
Synthesis of adenoviral targeting molecules by intein-mediated protein ligation
O Nyanguile1, C Dancik, J Blakemore
1Genetic Therapy, Inc., A Novartis Company, Gaithersburg, MD 20878, USA.
Abstract:
Adenoviral vectors infect cells through the binding of capsid proteins to cell-surface receptors. The ubiquitous expression of adenoviral receptors in human tissues represents an obstacle toward the development of systemically deliverable vectors for cancer therapy, since effective therapy may require delivery to specific sites. For these reasons, major efforts are directed toward the elimination of the native tropism combined with identification of ligands that bind to tumor-specific cell-surface proteins. Highthroughput technologies have identified potential targeting ligands, which need to be evaluated for their ability to retarget adenovirus to alternative receptors. Here, we present a strategy that permits the routine analysis of adenoviral targeting ligands. We use intein-mediated protein ligation as a means to produce functional biological molecules, that is, adenoviral targeting molecules that function as adapters between cellular receptors and the adenovirus fiber protein. We demonstrate the versatility of the present system by conjugating targeting ligands that differ in size and nature including an apolipoprotein E synthetic peptide, the basic fibroblast growth factor and folic acid. The resulting adenoviral targeting molecules mediate adenoviral gene delivery in cells that express the corresponding receptor.
Insights
Researchers developed a new method using intein-mediated protein ligation to create adenoviral targeting molecules. This strategy allows for routine analysis of ligands, enabling targeted gene delivery for potential cancer therapies.
Area of Science:
- Biotechnology
- Molecular Biology
- Gene Therapy
Background:
- Adenoviral vectors rely on capsid proteins binding to cell receptors for infection.
- Ubiquitous receptor expression hinders targeted cancer therapy delivery.
- Eliminating native tropism and identifying tumor-specific ligands are crucial for effective vector retargeting.
Purpose of the Study:
- To establish a versatile strategy for routine analysis of adenoviral targeting ligands.
- To create functional adenoviral targeting molecules that act as adapters between cellular receptors and the adenovirus fiber protein.
- To demonstrate the system's ability to retarget adenoviruses using various ligands.
Main Methods:
- Intein-mediated protein ligation was employed to synthesize adenoviral targeting molecules.
- Conjugation of diverse targeting ligands, including peptides and growth factors, to the adenovirus fiber protein.
- Evaluation of the retargeted adenoviral vectors' gene delivery efficiency in cells expressing specific receptors.
Main Results:
- A robust system for producing and analyzing adenoviral targeting ligands was established.
- Successful conjugation of apolipoprotein E peptide, basic fibroblast growth factor, and folic acid demonstrated system versatility.
- The resulting adenoviral targeting molecules effectively mediated gene delivery to cells expressing cognate receptors.
Conclusions:
- The developed intein-mediated ligation strategy provides a routine method for creating and evaluating adenoviral targeting molecules.
- This approach facilitates the development of systemically deliverable adenoviral vectors for targeted cancer gene therapy.
- The system's adaptability allows for the incorporation of various ligands to achieve specific cellular targeting.
