Viral particles with exogenous internal epitopes
A virion, exogenous technology used in the field of expression of peptides in viral capsids
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0159] This example illustrates the expression of peptides on the inner surface of viral capsids. In particular, a peptide derived from the mimotope of the group a determinant of HBsAg was inserted into the VP-S of CPMV. The sequence from an anti-idiotypic monoclonal antibody against hepatitis B virus surface antigen (HbsAg), AVYYCTRGYHGSSLY (SEQ ID NO: 33), and a highly hydrophobic octamer from the same sequence (GYHGSSLY; SEQ ID NO: 34) were reported It is effective in eliciting a homogeneous immune response using an "a" group determinant of the HBsAg itself (see, eg, US Patent Nos. 5,531,990; 5,744,135 and 5,856,087, which are incorporated herein by reference). The same peptides generate HBV-specific T helper cell responses. Therefore, it is called 2F10 and its derivatives are the analog sites of group a determinants. Pentamers and octamers from 2F10 can be expressed on the outer surface of CPMV particles. The octamer is expressed in the βBβC-loop of VP-S and the βEβA-lo...
Embodiment 2
[0162]This example illustrates the expression and internal display of a hydrophobic peptide corresponding to the CTL-epitope (MAL 7) of the circumsporozoite protein from Plasmodium berghei. P. berghei is a unicellular protozoan causative agent of human malaria. With respect to a peptide derived from an epitope of the circumsporozoite protein, having the amino acid sequence of SYIPSAEKI, can be expressed in the βBβC-loop of VP-S (producing a chimeric virion called Mal 4), and in the βBβC-loop of VP-S, and in the Two different sites at the carboxy-terminus are expressed (generating Mal 5 and Mal 6 in turn). The same peptide could be expressed at the amino terminal of VP-S, between repeats of the tyrosine residue at position 11 in the protein (Tyr11; Reference Example 1). The build is called MAL 7. The construct did not produce systemic symptoms at the initial stage after inoculation of the cDNA encoding the modified CPMV MAL 7 into the leaves of host cowpea plants. However, a...
Embodiment 3
[0164] This example demonstrates the in vivo generation and subsequent isolation of global mutations in chimeric viral genomes that confer selection on the infection progression and systemic spread of CVP internally expressing the MAL 7 peptide over homologous wild-type chimeric viral genomes sexual advantage. Surprisingly, the mutant chimeras described in Example 2 (above) were viable at the initiation and execution of a "natural" infection cycle by the recombinant virus, including systemic transmission in the host plant. However, in an effort to remove any structural constraints expected by selective pressure, the insertion site could have been altered in the mutations reported in MAL 7 described above. Examining the temperature factor of the crystal structure revealed that Val 10 is in a relatively rigid configuration, whereas residue Asp9 is very flexible. This implies that Val10Tyr11 may be repeated in order to transfer potentially destabilizing charged residues within t...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More