Team:Tsinghua/project

BACKGROUND

Monoclonal antibodies (mAb or moAb) are monospecific antibodies that are the same, made by identical immune cells that are all clones of a unique parent cell. When stimulated by almost any type of antigen, the immune system can create the specific antibody. This lays the foundation for monoclonal artificial antibodies.

The current most well-developed technique in Artificial Monoclonal Antibody is the famous Hybridoma Cell Production. Monoclonal antibodies are typically made by fusing myeloma cells with the spleen cells from a mouse that has been immunized with the desired antigen. The success rate is so low that a selective medium in which only fused cells can grow is used. This mixture of cells is then diluted and clones are grown from single parent cells on microtitre wells. The antibodies secreted by the different clones are then assayed for their ability to bind to the antigen. The most productive and stable clone is then selected for future use.

Project Outline

Tsinghua iGEM 2010 Project is focused on developing a new Antibody Production Method using Synthetic Biology. The Aim is to Simulate Antibody Selection and Production Technology with Bacteria. Traditional method of antibody production such as Hybridoma Technology is expensive and time consuming. In Hybridoma Technology, generation of B-Cell library is accomplished inside experimental animal, mouse for example; using Flow Cytometry to select a specific kind of antibody; then hybridoma would be prepared to harvest antibody. In this project, we expect to achieve all these processes cheap and efficiently in E.coli Cell. The whole project can be divided into two modules:

MODULE I: Generation of Antibody Library MODULE II: Selection of Specific Antibody

Module I

Abstract

Flow chart

Landing Pad Insertion

Helper Plasmid(HP) Insertion

Donor Plasmid(DP) Construction

Strategy 1

Strategy 2

Donor Plasmid(DP) Insertion & Recombination Induction

Removal of Helper Plasmid(HP)

Module II

Strategy 1

Strategy 2

Strategy 3

Cooperation with Macquarie_iGEM

Future