U.S. researchers have developed a set of guidelines for running plate-based chromatography experiments. The team, from the University of Virginia, developed the guidelines to help companies perform consistent, repeatable, and high-quality screening using 96-well plates.
Their aim is to help companies compare results between departments and across the wider industry, improving the quality of process development.
“Everyone runs different versions of these experiments and, depending on how they’re performed, you can get different answers to the same questions,” explains Nick Vecchiarello, PhD, assistant professor at the University of Virginia, who led the research. “We wanted to establish a more theoretical basis for how to do high-throughput screening, so that companies can get the results they expect.”
According to Vecchiarello, 96-well plate screening experiments are run to test how well commercial resins bind to proteins and impurities. The best-performing resin can then be used for product purification. Resins that bind too strongly make it hard to remove the protein, whereas resins with weak binding are poor at filtering out impurities. The new standards could help companies set criteria, such as limits on the strength of this binding, Vecchiarello explains.
Another issue in plate screening, he says, is that proteins are often strongly absorbed onto chromatography resin beads, meaning the remaining solution can be below the detection limits for experimental assays.
The new guidance, which was produced using a comprehensive academic literature search on plate-based experiments, could also help companies understand the detection threshold for their assays, he explains.
Supporting the guidance
To support the guidance, the team developed a software package that uses an Excel spreadsheet and a substantial body of academic literature to aid experimental design.
“We’re releasing the software package so that, if you’re a company or group within a company, it will help you design your experiments to meet the criteria you want,” says Vecchiarello, adding that historical data can also be fed into the software package.
The guidance and software are suitable for experiments on any protein-based product, including monoclonal antibodies, antibody drug conjugates, and potentially adeno-associated viruses.
The team says they’re now using the new guidance to help them research new chromatography materials.
