MilliporeSigma

HIV-1 Fusion Assay.

Bio-protocol (2014-08-20)
Marielle Cavrois, Jason Neidleman, Warner C Greene
ABSTRACT

The HIV-1 fusion assay measures all steps in the HIV-1 life cycle up to and including viral fusion. It relies on the incorporation of a β-lactamase Vpr (BlaM-Vpr) protein chimera into the virion and the subsequent transfer of this chimera into the target cell by fusion (Figure 1). The transfer is monitored by the enzymatic cleavage of CCF2, a fluorescent dye substrate of β-lactamase, loaded into the target cells. Cleavage of the β-lactam ring in CCF2 by β-lactamase changes the fluorescence emission spectrum of the dye from green (520 nm) to blue (447 nm). This change reflects virion fusion and can be detected by flow cytometry (Figure 2).

MATERIALS
Product Number
Brand
Product Description

Millipore
Millipore® Steriflip® Vacuum Tube Top Filter, pore size 0.22 μm, PVDF membrane, funnel capacity 50 mL, pack of 25 ea