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Published on: May 6, 2015
Targeting of adenoviral vectors through a bispecific single-chain antibody
H J Haisma1, J Grill, D T Curiel
1Department of Medical Oncology, University Hospital Vrije Universiteit, Amsterdam, The Netherlands. hj.haisma@azvu.nl
Abstract:
Recombinant adenoviral vectors are attractive in the context of cancer gene therapy because they are capable of delivering genes to a wide variety of tissues. The utility of adenoviruses is limited by their lack of specificity and by the absence of the receptor(s) for these viruses on many tumor cells. Redirecting adenoviral vectors to tissue- or tumor-specific targets can be achieved by using bispecific conjugates produced by chemical linkage of an anti-adenovirus antibody (Ab) and a ligand or Ab directed toward a specific target. To avoid the limitations of chemical conjugates, molecular conjugates of anti-fiber knob and ligand have been proposed. We present here a novel strategy that allows the production of recombinant bispecific single-chain Abs directed at cell surface molecules. A construct was made that encodes a neutralizing anti-adenovirus fiber single-chain Fv (scFv) Ab (S11) fused to a scFv Ab (425) directed against the epidermal growth factor receptor. The fusion protein markedly enhanced the infection efficiency of adenoviral vectors in epidermal growth factor receptor-expressing cell lines. The bispecific scFv could be purified and concentrated after binding of its 6His tag to a nickel column without significant loss of activity. This approach should permit the production of high quantities of active bispecific scFv for in vivo use. The universal design of the construct allows rapid screening for relevant specific scFv directed at cell surface antigens that can be incorporated into adenoviral targeting strategies.
Insights
Researchers developed a novel bispecific antibody strategy to improve adenoviral vector targeting for cancer gene therapy. This approach enhances gene delivery efficiency to specific tumor cells expressing the epidermal growth factor receptor.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Gene Therapy
Background:
- Recombinant adenoviral vectors are promising for cancer gene therapy due to broad tissue tropism.
- Adenovirus utility is hindered by poor specificity and lack of tumor cell receptors.
- Current targeting strategies involve chemical conjugates, which have limitations.
Purpose of the Study:
- To develop a novel strategy for producing recombinant bispecific single-chain antibodies (scFvs) for targeted adenoviral delivery.
- To create a fusion protein combining an anti-adenovirus antibody with an anti-epidermal growth factor receptor (EGFR) antibody.
- To enhance adenoviral vector infection efficiency in EGFR-expressing tumor cells.
Main Methods:
- Constructed a gene encoding a neutralizing anti-adenovirus fiber scFv (S11) fused to an anti-EGFR scFv (425).
- Produced and purified the bispecific scFv fusion protein using a 6His tag and nickel column chromatography.
- Evaluated the fusion protein's ability to enhance adenoviral vector infection in EGFR-expressing cell lines.
Main Results:
- The bispecific scFv fusion protein significantly enhanced adenoviral vector infection efficiency in EGFR-expressing cells.
- Purification via nickel column resulted in high yields of active bispecific scFv with minimal activity loss.
- The universal construct design facilitates rapid screening for other cell surface targets.
Conclusions:
- This novel bispecific scFv strategy offers a robust method for targeting adenoviral vectors to specific cells.
- The approach allows for the production of large quantities of active bispecific scFv for potential in vivo applications.
- This technology provides a versatile platform for developing targeted cancer gene therapies.
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