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Team:Wageningen UR/Project
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==== Introduction ==== | ==== Introduction ==== | ||
| + | The focus for the Hepatitis B VLP will be on vaccine production as final application. The final goal is to obtain a modified and bricked VLP containing an epitope. As a proof of principle we will link GFP to the outside and check the GFP with ELISA. GFP will be attached on the outside in two different ways: By fusion to the monomer of the VLP and via the K/E coil system. | ||
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| + | '''Aims''': Attach epitopes to the outside of the Hepatitis B VLP in order to make vaccines for pathogens other than Hepatitis B. Giving a proof of principle with GFP. Compare two different ways of attaching the GFP to the outside of the VLP. | ||
==== Outside Modification of the Hepatitis B VLP ==== | ==== Outside Modification of the Hepatitis B VLP ==== | ||
Revision as of 09:56, 25 May 2012
Contents |
Project VLPlatform
This year, the Wageningen UR team 2012 will work on the modification of virus-like particles (VLPs) to make them interesting platforms for vaccine production and/or site specific drug delivery. VLPs are empty virus capsids, meaning that it does not contain any viral genome, proteins and epitopes, except for the coat proteins. Coat proteins of some viruses have shown the ability to self-assemble in absence of its viral genome and other viral proteins, and thus form VLPs.
Introduction
The plan of our project is to use these VLPs to create a universal platform on which vaccines can be created or with which drugs can be delivered. To facilitate this, we want to put anchors on the outside of the VLP to attach antigens and ligands to, and anchors on the inside to attach medicine to, which is then encapsulated by the VLP. The anchors that we want to use are charged coils, which are already being used to encapsulate GFP into Cowpea Chlorotic Mottle Virus (CCMV) VLPs. This technique relies on charged coil peptides (negatively charged E-coil and positively charged K-coil) that can form ion bonds together. We want to use this technique to obtain a universal attachment system with which we can attach all kinds of epitopes to the VLP.
We selected 3 viruses that we want to use to produce VLPs with universal attachment units on the inside and outside of the VLPs. We have selected them based on existing experience at Wageningen University, increasing the probability of success and their promising structure. Besides CCMV, we will use the Hepatitis B core antigen VLP for the production of vaccines. The Potato Leaf Roll (PoLeRo) virus will be used to yield a newly expressed VLP in E. coli, because of its promising structure with outside spikes that are easy to modify. All three VLPs will be expressed in E. coli and all three will need to be submitted in the same standardised form for the competition. Therefore we will use the same E. coli strain for all three tracks. This increases the efficiency of the whole projects, because growing conditions will be the same.
Part 1: Modifying the CCMV VLP for site specific drug delivery
Introduction
CCMV will be one of the VLPs that we will be working on. We will focus on this VLP to prove the concept of using it as a Drug Delivery System.
Aim: Producing CCMV VLPs with epitopes on the outside or packaging coils in the inside, with the possibility to combine both to deliver a bio Nano carrier, with ligands on the outside and “medicine” packed on the inside.
Outside modification of CCMV VLP
inside modification of CCMV VLP
Part 2: Modifying the Hepatitis B VLP for Vaccine Production
Introduction
The focus for the Hepatitis B VLP will be on vaccine production as final application. The final goal is to obtain a modified and bricked VLP containing an epitope. As a proof of principle we will link GFP to the outside and check the GFP with ELISA. GFP will be attached on the outside in two different ways: By fusion to the monomer of the VLP and via the K/E coil system.
Aims: Attach epitopes to the outside of the Hepatitis B VLP in order to make vaccines for pathogens other than Hepatitis B. Giving a proof of principle with GFP. Compare two different ways of attaching the GFP to the outside of the VLP.
Outside Modification of the Hepatitis B VLP
Inside Modification of the Hepatitis B VLP
Part 3: Obtaining the Polero VLP expressed by E. coli
introduction
The Polero (potato leaf roll virus) virus coat proteins can be used as building blocks to form VLP’s, which has never been done before in E. coli. Therefore we will focus on the use of Polero virus coat proteins to make self-assembling VLP’s.
