Fused-silica capillaries chemically derivatized with silane/poly(vinylpyrrolidone) (PVP) or dynamically modified with plexiglas [poly(methyl methacrylate)] were prepared and evaluated with regard to column stability and separation performance for capillary isoelectric focusing of standard proteins. The PVP coating showed the better stability and was good for at least 100 runs while the plexiglas coating started to deteriorate after about 30 runs. The time spent for the plexiglas coating is about 40 minutes while the PVP coating requires two days. The migration time reproducibility was better with the PVP capillary (RSD 0.7-1.6%, n = 5) compared to the plexiglas-coated column (RSD 1.2-2.9%, n = 5) while peak area and height varied over a similar interval (RSD 2-28.1% area; 0.9-22.7% height, n = 5). The two most consistent proteins in this evaluation, viz. myoglobin A and carbonic anhydrase II, showed linear dynamic ranges between 5-150 and 5-50 microg/mL, and limits of detections at 2 and 1 microg/mL, respectively, employing UV detection at 280 nm.
Four years ago, the first acoustic droplet ejectors (ADEs) were launched on the market, providing a new generation of high-throughput noncontact liquid handlers that outclassed traditional contact instruments in almost every respect. This introduction of noncontact dispensing has triggered radical changes to the screening/compound management interface. Higher quality is achieved through greater accuracy and precision, whereas lower sample volumes can be used, and 1536 plate formats have become a reliable reality. Prior to the ADE instrument launch, 1536 assay-ready plate preparation was a high-effort enterprise requiring users to spend time developing liquid-handling methods along with daily fine-tuning of instruments to reach the desired level of performance. By overcoming the nanoliter dispensing hurdle and successfully transferring assays to high-density formats, a new dimension for cutting costs has emerged. Once the screening customer has adapted to this new world, the rules of supply can also change, with the traditional automated plate store no longer being necessary when the compound library can be stored in 1536 plates. Processing efficiency recently has been further supported by innovative new automation-friendly solutions such as plate desealers, prolonging the life span of working plate copies. Both cost and waste control have never had a higher profile, and noncontact dispensing contributes to these important areas. In some processes (e.g., when piercing septa), contact dispensing remains the best option, but cost control is still essential, and an innovative solution to minimize DMSO consumption from tip washing has had a big impact on consumable budget without compromising quality.
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