Problem
The synthesis of vitamin D is dependent on the exposure of skin to UV light from sunlight. The characteristics of northern climates, including cold weather and limited sunlight, pose a danger concerning vitamin D deficiencies. In Canada, 33% of the population experiences insufficiency1. Worldwide, it is estimated that around 1 billion people experience low vitamin D levels2. In northern communities in British Columbia, Canada, vitamin D testing is expensive and time-consuming; local laboratories do not have the necessary equipment, and so blood samples must be transported to the lower mainland for testing.
Goals
The challenges posed by current vitamin D testing methods in northern BC illustrate the need for an inexpensive, rapid, and simple vitamin D test.
Inexpensive - Our system is more cost-effective than current techniques in our region, which require specialized equipment that necessitates transport of samples.
Rapid - The generation of a fluorescent signal by our strain’s molecular machinery will allow for efficient quantification of an individual’s vitamin D levels.
Simple - Our plans for a benchtop kit ensure that the test can be performed easily, with minimal equipment requirements.
System
Purify - Existing methods are used to purify the inactive form of vitamin D (25(OH)D) from a blood sample. The freed vitamin D diffuses into our specialized strain of E. coli.
Activate - Recombinant 1α-hydroxylase activates the 25(OH)D, yielding 1,25(OH)2D.
Detect - 1,25(OH)2D binds to the ligand-binding domain of vitamin D receptor protein, which is coupled to a FRET system. The binding of vitamin D causes the fluorophores to move closer, allowing for a measurable signal.
References
1. https://www150.statcan.gc.ca/n1/pub/82-624-x/2013001/article/11727-eng.htm
2. Palacios, C., & Gonzalez, L. (2014). Is vitamin D deficiency a major global public health problem?. The Journal of steroid biochemistry and molecular biology, 144 Pt A, 138–145. https://doi.org/10.1016/j.jsbmb.2013.11.003