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Published on: August 14, 2011
Trastuzumab-binding peptide display by Tobacco mosaic virus
Olga Y Frolova1, Igor V Petrunia, Tatiana V Komarova
1A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University and N.I. Vavilov Institute of General Genetics, Russian Academy of Science, Moscow 119992, Russia.
Abstract:
Human epidermal growth factor receptor-2 (HER2/neu) is a target for the humanized monoclonal antibody trastuzumab. Recently, trastuzumab-binding peptides (TBP) of HER2/neu that inhibit proliferation of breast cancer cells were identified. We have now studied conditions of efficient assembly in vivo of Tobacco mosaic virus (TMV)-based particles displaying TBP on its surface. The system is based on an Agrobacterium-mediated co-delivery of binary vectors encoding TMV RNA and coat protein (CP) with TBP in its C-terminal extension into plant leaves. We show how the fusion of amino acid substituted TBP (sTBP) to CP via a flexible peptide linker can improve the manufacturability of recombinant TMV (rTMV). We also reveal that rTMV particles with exposed sTBP retained trastuzumab-binding capacity but lost an anti-HER2/neu immunogenic scaffold function. Mouse antibodies against rTMV did not recognize HER2/neu on surface of human SK-BR-3 cells.
Insights
Researchers developed Tobacco mosaic virus (TMV) particles displaying trastuzumab-binding peptides (TBP) to target HER2/neu in breast cancer. These engineered TMV particles bind trastuzumab but lose immunogenic properties.
Area of Science:
- Plant virology
- Molecular biology
- Cancer research
Background:
- Human epidermal growth factor receptor-2 (HER2/neu) is a key target for breast cancer therapy, notably with the monoclonal antibody trastuzumab.
- Trastuzumab-binding peptides (TBP) derived from HER2/neu have been identified for their potential to inhibit cancer cell proliferation.
- Tobacco mosaic virus (TMV) offers a platform for displaying peptides on its surface for potential therapeutic applications.
Purpose of the Study:
- To investigate efficient in vivo assembly of TMV-based particles displaying TBP.
- To evaluate the manufacturability and functional characteristics of recombinant TMV (rTMV) displaying modified TBP (sTBP).
- To assess the trastuzumab-binding capacity and immunogenic properties of the engineered rTMV particles.
Main Methods:
- Agrobacterium-mediated co-delivery of binary vectors encoding TMV RNA and coat protein (CP) fused with TBP into plant leaves.
- Engineering of rTMV by fusing amino acid substituted TBP (sTBP) to CP via a flexible peptide linker.
- Assessing trastuzumab-binding capacity of rTMV particles displaying sTBP.
- Evaluating the immunogenic scaffold function and HER2/neu recognition by mouse antibodies against rTMV.
Main Results:
- Fusion of sTBP to CP via a flexible linker improved the manufacturability of rTMV.
- The engineered rTMV particles with exposed sTBP retained their capacity to bind trastuzumab.
- The rTMV particles lost their anti-HER2/neu immunogenic scaffold function, as evidenced by mouse antibodies not recognizing HER2/neu on SK-BR-3 cells.
Conclusions:
- Recombinant TMV particles can be efficiently assembled in vivo to display TBP, with improved manufacturability achieved through sTBP fusion to CP.
- These engineered TMV particles maintain trastuzumab-binding activity, suggesting potential for targeted delivery or diagnostics.
- The loss of immunogenic scaffold function indicates a dissociation between peptide display and immune response induction, important for therapeutic design.
