Revvity, 64AAV5PEH, HTRF AAV5 Detection Kit, 10,000 Assay Points

Overview
Adeno-associated virus (AA) vectors are the leading platform for gene delivery for the treatment of a variety of human diseases. AAV Serotype 5 (AAV5) has been consistently characterized as the serotype with the lowest seroprevalence and minimal cross reactivity against pre-existing neutralizing antibodies which can be an advantage for its use as a vector for gene therapy of hemophilia A and diseases associated with its natural tissue tropism such as lung, retina, or central nervous system cells. The AAV5 kit is designed to detect and quantify AAV5 particles in an easy-to-use, no-wash format. The simple and robust procedure benefits from increased throughput compared to ELISA.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
AAV5 Capsid assay principle
The Adeno-Associated Virus serotype 5 (AAV5) assay measures AAV5 capsid in cell supernatant or cell lysate. The assay uses two antibodies anti-AAV5: one labelled with d2 (acceptor) and the other to biotin that binds to Streptavidin Eu-cryptate (donor) in a premix. In presence of AAV5 capsid in a cell extract or supernatant the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor, and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the capsid present in the sample and provides a means of assessing any changes caused by experimental variability under a no-wash assay format.

 

AAV5 capsid assay protocol
The AAV5 capsid assay protocol, using a 384-well small volume white plate, is described on the right. 5 µL of sample or standard and 5 µL of diluent are dispensed directly into the detection plate for detection by HTRF® reagents. The Biotin antibody anti-AAV5 was pre-mixed with Streptavidin labeled with donor before being added. The assay can be run in up to a 1536-well format by simply resizing each addition volume proportionally. Application-Bioprocessing
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4
Product Group-Kit
Sample Volume-5 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Viral Particles
Technology-TR-FRET
Unit Size-10,000 assay points