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Dalvano and Kulp Lab: High Throughput Drug Screening

Principal Investigators: Brielle Dalvano, Research Fellow, and John Kulp, PhD, Computational Chemistry

The Kulp lab is interested in finding economic and robust methods to screen large numbers of drug candidates. Traditional drug screening is done in 96-well plates. The Kulp lab routinely runs 384-well assays and is beginning to use 1,536 well plates. Yes, that is 1,536 reactions on one small plate! The statistics and scientific design become increasingly important in these high-throughput assays. We are applying the screening technologies to the following therapeutics areas:

A. High-throughput polymerase chain reaction assay for the detection of viral DNA

Hepatitis B is a double-stranded DNA virus that is excreted from infected cells in virion particles. These virion particles encapsulate viral DNA in two membranes, including an outer envelope membrane containing hepatitis B virus (HBV) surface proteins and a nucleocapsid of HBV core proteins. The extraction of viral DNA for detection in high-throughput drug screening assays is complicated by the DNA’s outer membrane and nucleocapsid encapsulation. We are developing a method that allows for simple extraction of viral hepatitis B DNA from virion particles and subsequent detection using polymerase chain reaction (PCR). The method aims to allow for a novel assay for PCR based high-throughput drug screening of potential drug candidates to treat HBV that reduces the typically high costs associated with PCR and HBV DNA extraction.    

 

B. Enzyme linked immunosorbent assay (ELISA) for the detection of surface antigen from the hepatitis B virus (HBsAg)

ELISA assays are one of the most common assays run at large pharmaceutical companies because of their high sensitivity and specificity. We are running ELISA assays in 384 well-plates looking for drug like small molecule inhibitors of a viral protein that leads to T cells exhaustion and dampens the immune system allowing the hepatitis B virus to be tolerated. Finding a functional cure for hepatitis B will likely require an immune system adjuvant that will either be host targeting or likely target a direct viral product like the S antigen.

 

Additional Information:

  1. Bleicher, K. H.; Bohm, H. J.; Muller, K.; Alanine, A. I., Hit and lead generation: beyond high-throughput screening. Nat Rev Drug Discov 2003, 2 (5), 369-78.
  2. Gadkar, V.; Filion, M., New Developments in Quantitative Real-time Polymerase Chain Reaction Technology. Curr Issues Mol Biol 2014, 16, 1-6.
  3. Gan, S. D.; Patel, K. R., Enzyme immunoassay and enzyme-linked immunosorbent assay. J Invest Dermatol 2013, 133 (9), e12.
  4. Lorenz, T. C., Polymerase chain reaction: basic protocol plus troubleshooting and optimization strategies. J Vis Exp 2012, (63), e3998.