Genetic Fusion-based VLP Modification Service
As is a leading global custom service provider, CD BioSciences dedicated to delivering highly customized and integrated solutions with the best quality to advance our customers' projects. CD BioSciences has a sophisticated VLPlantTM platform and extensive experience dedicated to providing our customers with genetic fusion methods to modify virus-like particles (VLPs).
Overview of Genetic Fusion-based VLP Modification
Genetic fusion mainly refers to the use of biochemical methods to modify DNA sequences, introduce target gene fragments into host cells, or delete specific gene fragments from the genome, thereby changing the host cell genotype or making the original genotype enhanced. Genetic fusion has been widely used in all areas of human life, and genetic fusion of VLPs involves the fusion of genes of the desired surface ligands with genes of the outer shell proteins of VLPs by covalent attachment. Although the added ligand (peptide/protein) may inhibit protein folding and proper assembly of VLPs, this approach is applicable to the generation of most cVLPs.
Characteristics of the Genetic Fusion
- More stable binding of VLPs to antigens
- Simple manufacturing process
- Only protein-based antigens can be linked to VLPs by gene fusion to form cVLPs
- The outcome of gene fusion is unpredictable and depends on many factors including the length and purity of the target antigen peptide
- Usually only peptides shorter than 30 amino acids will not interfere with the correct assembly of VLPs
- The presence of heterologous antigens can lead to improper folding of VLPs or the formation of cVLPs of varying sizes
The use of gene fusion to fuse exogenous peptides to the intact or truncated N-terminus/C-terminus (N/C-ter) is the most appropriate strategy for the design and construction of cVLPs. This approach usually does not interfere with the primary structure of the VLPs and does not affect the immunogenicity of the resulting cVLPs. In some cases, truncating the N/C-ter of Cap not only does not affect the assembly of VLPs, but also facilitates their expression and assembly into cVLPs. This strategy often uses specific tags, such as His tags to aid purification and GFP tags for intracellular localization. Therefore, the conformational position of the two N/C-ter of Cap (embedded in the interior of the cVLPs or facing its surface) generally determines the feasibility and efficiency of this strategy.
3D structure of four representative cVLPs. (Lei X, et al., 2020)
CD BioSciences offers services covering all experimental elements related to the construction of cVLPs using genetic fusion methods for plant platforms. If you are interested in our services, you can contact us for more details.
Workflow of Our Service
- Clone the target sequence
- The target sequences were screened for codon optimization.
- Design specific primers.
- The target sequence was cloned by PCR between the promoter and terminator (whose repressor sequence was modified at the 5' end).
- Construction of chimeric structures
- Screen and synthesize antigenic epitopes.
- The N/C-terminal amino acids of the target sequence were replaced with the antigenic epitope by PCR.
- The resulting chimera structure was verified by sequencing.
- Construction of expression vectors
- The chimeric structures were then cloned into expression vectors and used in plant platforms to construct VLPs.
- Production of VLPs
- The constructed vectors were transferred into Agrobacterium using electroporation transformation to produce cVLPs.
- Lei, X.; et al., Genetic engineering strategies for construction of multivalent chimeric VLPs vaccines. Expert Rev Vaccines. 2020, 19:235-246.