Team:Queens Canada/Laboratory-Overview

Overview

Overview

The goal of this year’s project is to develop a novel antibody-based test to detect the bacteria that causes Lyme Disease (Borrelia Burdorferi), through outer surface protein A (OspA). To develop this test, we created a single chain variable-fragment (ScFv) to express in an E. coli system, based on previous research by Ghosh and Huber (1). An ScFv was selected for expression in E. coli as these structures do not require the post-translational modifications essential for the creation of full-size IgG antibodies and thus can be expressed by common bacteria. The ScFv structure was the foundation of our modular protein which we then linked to both an Alkaline Phosphatase and a Green Fluorescence Protein to aid in binding detection and protein folding confirmation, respectively.

Borrelia Burdorferi
Depiction of Borrelia Burgdorferi spirochetes

Figure 1 - IgG antibody labeled schematic and an scFv graphic with ospA labelled ligand
A graphic showing the various components of an IgG antibody and the single chain variable fragment (scFv) component of the antibody which we are using in our bioreceptor/biosensor system. The ligand to the scFv, ospA, is the primary target protein present on the surface of Borrelia burgdorferi cells used for detection.

References

1. Ghosh, S., and Huber, B. T. (2007) Clonal diversification in OspA-specific antibodies from peripheral circulation of a chronic Lyme arthritis patient. J. Immunol. Methods. 321, 121–134


Explore these areas below.


Design

Before we could get into the lab, we had to make sure we had done some groundwork first to make sure we were on the right track with our experiments.

Protocals

Documentation describing protocols and procedures followed in the lab during experiments.

Experiments

Get an inside look at the experiments we conducted while creating our biosensor, including PCR amplification, digestion and ligation, cell culturing, protein expression, SDS page, and many more!

Results

Here is where we summarize the outcome of all our laboratory work conducted this summer.

Safety

A breakdown of all the of the Wet Lab safety training we conducted to ensure out team was safe throughout the summer.


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