Revvity, 64AAV3PEH, HTRF AAV3B 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 3B (AAV3B), when used as vector in gene therapy, has a liver cell transduction efficiency greater than those of other serotypes which has resulted in efforts to develop this virus as a gene therapy vector for hemophilia. AAV3B also exhibits a natural tropism for other tissues such as brain, muscle, and retina. The AAV3B kit is designed to detect and quantify AAV3Bparticles 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
AAV3B capsid assay principle
The Adeno-Associated Virus serotype 3B (AAV3B) assay measures AAV3B capsid in cell supernatant or cell lysate. The assay uses two anti-AAV3B antibodies: one coupled to HRP that binds to anti-HRP d2 (acceptor) in premix 1, and the other coupled to biotin, that binds to Streptavidin Eu-cryptate (donor) in premix 2. In presence of AAV3B 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.

 

AAV3 Capsid assay protocol
The AAV3B 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 plate for detection by HTRF® reagents. The Biotin anti-AAV3B antibody is pre-mixed with Streptavidin labeled with the donor, and HRP anti-AAV3B is pre-mixed with anti-HRP labeled with the acceptor. 5 µL of each premix are 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
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