AI Article Synopsis
- HIV-1 infection starts when the gp120 protein binds to the CD4 receptor on cells, leading to changes that allow it to also bind to the CCR5 or CXCR4 coreceptors.
- Researchers have created a full-length single chain protein (FLSC) that mimics these changes in gp120, which they used to develop two types of assays: flow cytometry and ELISA.
- The ELISA-based assay proved to be more effective and efficient for identifying inhibitors of gp120 binding to CCR5, showing better sensitivity and suitability for high throughput screening compared to the flow cytometry method.
Article Abstract
HIV-1 infection is initiated by the interaction of the envelope glycoprotein gp120 with the cellular receptor CD4 that triggers conformational changes in gp120 necessary for subsequent interaction with a coreceptor CCR5 (or CXCR4). The CD4-induced (CD4i) conformation of gp120 can be mimicked by a full-length single chain (FLSC) protein consisting of gp120 linked with the D1D2 domains of CD4 by a 20-amino-acid linker. We have used this protein to establish a flow cytometry-based assay and an ELISA-based assay to identify inhibitors that block the binding of gp120 to CCR5. Both assays are specific for detecting the known CCR5 antagonist TAK-779, but the ELISA-based assay was more sensitive, simple, inexpensive, and rapid; thus, it can be adapted to high throughput screening (HTS). The ELISA-based method was validated with a diverse set of known antagonists, for example, TAK-779, AOP-RANTES, PSC-RANTES, and several mAbs.
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| http://dx.doi.org/10.1016/j.virol.2004.06.022 | DOI Listing |
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