Technical library

Simple Western provides new vaccine technology to combat outbreaks. Simple Western instruments outperform traditional Western blot at every stage of the vaccine development pipeline.

Over the course of Dr. Nicolas Bazan’s several decades-long career, he has been a research scientist, teacher, mentor, community leader, author, screenplay co-writer, executive movie producer, patron of the arts, and entrepreneur. Dr. Bazan, who has published close to 500 peer-reviewed publications, sings the praises of Simple Western assays with Jess. Jess has dramatically improved the protein analytical methods on which his laboratory relies to study the cellular and molecular events of neurodegenerative disease onset and progression.

Adeno-associated viruses (AAV) are promising vectors for the delivery of genetic material in gene therapy. During the manufacture of AAV, critical quality attributes like charge heterogeneity and purity must be carefully monitored because they can impact the product’s safety and efficacy. Imaged capillary isoelectric focusing (icIEF) and capillary electrophoresis sodium dodecyl sulfate (CE-SDS) are two powerful methods to respectively characterize charge heterogeneity and purity, but traditionally two separate platforms are required to run these methods. Here, we used a single platform to develop icIEF and CE-SDS methods to analyze AAV2 and AAV6 serotypes to monitor product stability, identity and purity. We show that these methods could reproducibly quantify both intact (by icIEF) and denatured AAV (by icIEF and CE-SDS) samples. The CE-SDS method could separate and quantify individual AAV capsid proteins, showed robust repeatability (<5% RSD) while also detecting impurities. The icIEF method was useful for measuring both denatured and intact particles with high repeatability (<4% RSD). Interestingly, preliminary evidence suggests that icIEF can also distinguish between full (1 x 10^13 GC/mL) and ‘empty’ (<10^12 GC/mL) AAV capsids.

To characterize the efficacy of Degrader molecules, researchers generally run dose response curves by way of traditional SDS-PAGE Western blotting methods. Simple Westerns let you separate and analyze proteins by size (or charge) from 2 kDa to 440 kDa in just 3 hours.

In this Application Note, we show you how it may be possible to optimize the signal of poorly behaving antibodies with the Signal Enhancement Reagent on Milo.