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Matsuyama-shi, Japan

Matsunaga S.,Yokohama City University | Masaoka T.,Clinical Research Center | Sawasaki T.,Ehime University | Morishita R.,Yokohama City University | And 10 more authors.
Frontiers in Microbiology | Year: 2015

Due to their high frequency of genomic mutations, human retroviruses often develop resistance to antiretroviral drugs. The emergence of drug-resistant human immunodeficiency virus type 1 (HIV-1) is a significant obstacle to the effective long-term treatment of HIV infection. The development of a rapid and versatile drug-susceptibility assay would enable acquisition of phenotypic information and facilitate determination of the appropriate choice of antiretroviral agents. In this study, we developed a novel in vitro method, termed the Cell-free drug susceptibility assay (CFDSA), for monitoring phenotypic information regarding the drug resistance of HIV-1 protease (PR). The CFDSA utilizes a wheat germ cell-free protein production system to synthesize enzymatically active HIV-1 PRs directly from PCR products amplified from HIV-1 molecular clones or clinical isolates in a rapid one-step procedure. Enzymatic activity of PRs can be readily measured by AlphaScreen (Amplified Luminescent Proximity Homogeneous Assay Screen) in the presence or absence of clinically used protease inhibitors (PIs). CFDSA measurement of drug resistance was based on the fold resistance to the half-maximal inhibitory concentration (IC50) of various PIs. The CFDSA could serve as a non-infectious, rapid, accessible, and reliable alternative to infectious cell-based phenotypic assays for evaluation of PI-resistant HIV-1. © 2015 Matsunaga, Masaoka, Sawasaki, Morishita, Iwatani, Tatsumi, Endo, Yamamoto, Sugiura and Ryo. Source


Takemori N.,Ehime University | Takemori A.,Ehime University | Matsuoka K.,Ehime University | Morishita R.,CellFree science Co. | And 6 more authors.
Molecular BioSystems | Year: 2015

Using a wheat germ cell-free protein synthesis system, we developed a high-throughput method for the synthesis of stable isotope-labeled full-length transmembrane proteins as proteoliposomes to mimic the in vivo environment, and we successfully constructed an internal standard library for targeted transmembrane proteomics by using mass spectrometry. © The Royal Society of Chemistry 2015. Source


Trademark
CellFree science Co. | Date: 2006-05-23

Chemical reagents for research purposes.


Trademark
CellFree science Co. | Date: 2006-04-04

Automatic protein synthesizer.


Trademark
CellFree science Co. | Date: 2006-05-23

Chemical reagents for research purposes.

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