News Gator Flex SA Kit – A Reactivable Streptavidin Probe Kit Introduction Many of the label-free biosensors have been regenerated successfully to reduce cost for drug research and discovery. However, extensive application of streptavidin biosensors is still hindered by the irreversible immobilization of the bait molecule, and the ability to use the same probe for different application is desirable for cost effective research. Gator Flex SA Kit Gator® Bio’s Gator Flex SA Kit is the first in the market to provide reactivable streptavidin probes with a reagent set. One probe can easily be used by different users with different applications, which makes it very useful for academics or for use in fully automated screening of different biomolecular interactions. The Gator™ platform enables researcher to monitor whole reactivation process rapidly and provides flexibility and confidence. Performance Summary Remove both capture reagent and biotin-bait molecule and immobilize new capture reagent < 5 minutes Use with same or different biotinylated bait Can be re-used 10 times or more without loss of performance Automated reactivation Reagents can be stored at 4°C for up to 5 months Results Loading height reproducibility Three Flex SA probes were tested over 10 reactivations using biotinylated TNF-α at 1µg/mL. The table below shows CV < 5.0% suggesting excellent loading height reproducibility. Reproducibility of KD over 10 reactivations using the same bait Kinetics was evaluated using biotinylated TNFα and anti-TNFα antibody using a 5 min 1000 rpm pre-wet in K buffer and capture of Flex SA Capture Reagent (SA-CR). The biotinylated TNF-α was loaded onto the sensor, then exposed to association and dissociation of 10, 30, 100 and 300 nM anti TNF-α antibody. Global-fit analysis using Gator Bio software for the TNF-α binding interaction with anti- TNF-α resulted in KD = 7.63E-10 M. KD measurements for different proteins over 10 reactivations The same probe was tested with 2 different proteins over 10 reactivations. A CV < 5% was observed for both proteins. Gator Bio Total Solution GatorTM Flex SA kit GatorTM Prime or GatorTM Plus Gator Bio Software Ordering info PL168-350001: GatorTM Flex SA Kit
News Gator® AAVX Probes for Rapid and Label-free Quantitation of AAV Serotypes Introduction Adeno associated virus (AAV) capsids are a leading modality for in vivo gene delivery. Complete and precise characterization of capsid particles, including capsid and vector genome concentration, is necessary to safely and efficaciously dose patients. In virus development and production, it is important to determine the virus concentration at different stages of the process, to optimize the clone used as well as the production yields. Biolayer Interferometry (BLI) using AAV specific biosensors is a rapid label-free method for quantitation of AAV serotypes. Gator AAVX Probes The Gator AAVX probes are a high specificity antibody-based biosensors that enable direct capture and quantitation of different serotypes of AAV in crude lysates, column eluates, cell lysates and cell culture supernatants, serving as an alternative to traditional time-consuming analytical methods, such as qPCR, ddPCR, Dot blot and ELISA. The Gator AAVX probe uses proven CaptureSelect™ (Thermo Fisher Scientific) high affinity and high specificity anti-AAVX antibody. Performance Summary Dynamic range: 1 x 109–1 x 1013 vp/mL for most AAV serotypes Throughput: 8 samples in 10 minutes, 96 samples in 120 minutes Limit of detection: typically, 5 x 108 vp/mL (serotype dependent) Crude sample tolerant Stable over broad pH range Cost effective – Reusable at least 10 times by regeneration Results Dynamic range: The AAV2 serotype range was tested using the Gator AAVX probes. The data shows 4 orders of magnitude dynamic range. The data was acquired using 1:3 dilution in 10 min at 1,000rpm. Accuracy and reproducibility AAV5 serotype was used to test the accuracy and reproducibility. The table below shows recovery close to 100% and CV ranging from around 1.5% at medium and high titers to 10.9% at low titer. Binding rate of different AAV serotypes The binding rate of 8 different serotypes of AAV were studied. The figure below shows binding rates of tested serotypes at 2 x 1011 vp/mL concentration. Regeneration performance Data below shows performance after 10 regenerations of the same probe. No loss in binding rate observed even after 10 regenerations Gator Bio Total Solution GatorTM AAVX Probes GatorTM Prime OR GatorTM Plus Gator Bio Software Ordering info PL168-160017: GatorTM AAVX Probe