Filtration improves the performance of a high-throughput screen for anti-mycobacterial compounds.
The tendency for mycobacteria to aggregate poses a challenge for their use in microplate based assays. Good dispersions have been difficult to achieve in high-throughput screening (HTS) assays used in the search for novel antibacterial drugs to treat tuberculosis and other related diseases. Here we describe a method using filtration to overcome the problem of variability resulting from aggregation of mycobacteria. This method consistently yielded higher reproducibility and lower variability than conventional methods, such as settling under gravity and vortexing.
Center for Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Nancy
N
Cheng
Center for Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Melissa A
MA
Porter
DMPK Advisors, Chapel Hill, North Carolina, United States of America.
Lloyd W
LW
Frick
Center for Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Yvonne
Y
Nguyen
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Jennifer D
JD
Hayden
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Ellen F
EF
Young
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Miriam S
MS
Braunstein
Center for Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina, United States of America.
Emily A
EA
Hull-Ryde
Center for Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina, United States of America; Cancer Genetics Program, Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina, United States of America.
William P
WP
Janzen
D000995:Antitubercular Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005374:Filtration / Q000295:instrumentation / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods; D008851:Micropore Filters; D020102:Mycobacterium smegmatis / Q000187:drug effects* / Q000502:physiology; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000502:physiology; D015203:Reproducibility of Results
D000995:Antitubercular Agents