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


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