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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
An in vitro caseum binding assay that predicts drug penetration in tuberculosis lesions
Journal of Visualized Experiments, Volume 2017, No. 123, Article e55559, Year 2017
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Description
The eradication of tuberculosis disease requires drug regimens that can penetrate the multiple layers of complex pulmonary lesions. Drug distribution in the caseous cores of cavities and lesions is especially crucial because they harbor subpopulations of drug-tolerant bacteria also commonly referred to as persisters. Existing methods for the measurement of drug penetration in tuberculosis lesions involve costly and timeconsuming in vivo pharmacokinetic studies coupled to bioanalytical or imaging techniques. The in vitro measurement of drug binding to caseum macromolecules was proposed as an alternative to such techniques since this binding hinders the passive diffusion of drug molecules through caseum. Rapid equilibrium dialysis is a fast and reliable system for performing plasma protein and tissue binding studies. In this protocol, we used a rapid equilibrium dialysis (RED) device to measure drug binding to homogenates of caseum that is excised from the lesions and cavities of tuberculosis-infected rabbits. The protocol also describes how to generate a surrogate matrix from lipid loaded THP-1 macrophages to use in place of caseum. This caseum/surrogate binding assay is an important tool in tuberculosis drug discovery and can be adapted to help study drug distribution in lesions or abscesses caused by other diseases. © 2017 Journal of Visualized Experiments.
Authors & Co-Authors
Sarathy, Jansy Passiflora
United States, Newark
Public Health Research Institute
Weiner, Danielle M.
United States, Bethesda
National Institutes of Health Nih
Gonzales, Jacqueline
United States, Bethesda
National Institutes of Health Nih
Via, L. E.
United States, Bethesda
National Institutes of Health Nih
Dartois, Véronique A.
United States, Newark
Public Health Research Institute
Statistics
Citations: 17
Authors: 5
Affiliations: 2
Identifiers
Doi:
10.3791/55559
ISSN:
1940087X
Research Areas
Noncommunicable Diseases