Automation using IMCStips® with
Hamilton Microlab® STAR™
Presenters and Moderator
Presenter: Yuko Ogata, PhD
Moderator: Richard S. Rogers, PhD
Executive Summary
Manual sample preparation is not suited for a large set of samples, is amenable to human errors, and creates challenges when implementing identical processes across multiple locations. In addition, sample recovery from size-exclusion spin columns can vary greatly depending on the initial sample concentrations. The automated sample preparation using IMCStips provides comparable relative PQA values to manual preparation with excellent reproducibility, improved recoveries across different concentrations, more streamlined walk away solution, and higher throughput capability. The seamless sample preparation without intervention from the start to finish makes the MAM more suitable for all stages of drug development and production.
Challenges
There is a growing need for sequence confirmation and attribute analytics by MAM for a large set of samples. This platform is new and evolving and the company foresees many samples (ex. 96 samples at a time) requiring MAM analysis soon as it is no longer practical to prepare samples manually. In manual sample preparation workflow, dilution and digestion are not suitable for all samples with trypsin (incomplete digestion depending on the molecules, starting sample concentration, or enzymes); the percent sample recovery suffers for low concentration samples (<2 mg/mL) upon size exclusion chromatography (SEC) buffer exchange; and the method is susceptible to person-to-person variability.
How IMCS Helped
SizeX IMCStips helped to greatly enhance the MAM automation protocol. It resulted in a higher and more reliable percent recovery at buffer exchange, which allowed the company to omit concentration measurements (A280) after buffer exchange. Analysis showed consistent mass spectrometry TIC levels and PTM results within plates and over a wide range of concentrations. This organization process enhanced sample throughout while generating comparable results to manual preparation. As a result, the seamless sample preparation with intervention from start to finish makes the MAM more suitable for all stages of drug development and production.
Results
Quotes from Presentation
“It’s a great timesaving, especially for large set of samples. A small batch of manual sample prep takes about up to 3 hours or so, but I cannot do 96 samples at a time. So, by being able to do a large set of samples, it saves loads of time instead of doing manual rounds of sample prep. Once you set up the tips and reagents, it’s fairly hands off. You have much less bench time.”
"The scripts were made by IMCS, but of course it is customizable. We had great support from IMCS as well as Hamilton. During the initial set up, they tested the script and made sure that it’s comparable to the range of concentrations I will be proving. So, a great collaboration between IMCS and Hamilton and I really enjoyed the help from them.”
"The percent sample yields at buffer exchange with SizeX tips are significantly better for lower concentration samples. In other words, we have more concentrated samples during that digestion stage and after TSA cleanse.”
