From 2012.igem.org

Revision as of 09:57, 25 May 2012

• Home
• Team
  • The Team
  • Team Registration
• Project
  • 1. Introduction
  • 2. PnA System
  • 3. Virus-Like Particles
    • CCMV
    • HepB
    • Polerovirus Natural BioBrick
  • 4. Outside Modification
  • 5. Inside Modification
  • 6. Detection of VLPs
  • 7. Applications
  • 8. The Constructor
  • 9. Final Overview
• Human Practices
  • Human Practices Home
  • 1. Mini Symposium
  • 2. Visiting Secondary Schools
  • 3. Discovery Festival
    • Communication Science
  • 4. Stakeholders
  • 5. Munich CAS Conference
• Safety
  • Introduction
  • 1. General safety
  • 2. Virus-related safety
  • 3. Regulations
  • 4. Safety suggestions
  • 5. Safety of applications
• Modeling
  • Human Body Model
  • Tertiary folding prediction
  • VLP assembly model
• Attributions
• Parts
• Notebook
  • Journal
  • Protocols

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Last year's project: Synchronized Oscillatory System

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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.

Aim: Our primary aim is to get self-assembling VLP’s from the Polero virus in E. coli. If we succeed, we can attempt to modify the spike on the outside.

Results