Purification of HTOS Libraries by Supercritical Fluid Chromatography

Evaluation and Implementation of a Commercially Available Mass-Guided SFC Purification Platform in a High Throughput Purification Laboratory in Drug Discovery

“…[19,21,27] The use of additives is not favored in our high throughput laboratory, so samples requiring better peak shape than what is delivered by the 2-ethylpyridine column are analyzed with comparable gradients on the aminophenyl, nitro, and silica columns. The best alternative stationary phase is then utilized during purification of the difficult samples.…”

“…Two studies also demonstrate comparable recovery when samples were purified by both techniques. [17,19] The use of preparative SFC for high throughput purifications initially focused on UV based fraction triggering systems. [10,[20][21][22][23] The early work of Berger, Farrell, and White in this area clearly showed that the proposed advantages of the SFC approach could indeed be realized.…”

Advances in High Throughput Supercritical Fluid Chromatography

“…[61,67,68] Gradient elution is almost always necessary to achieve both separation and elution of the solutes. [58][59][60][61]68,69] However, if an adequate separation is not achieved, changing the stationary phase has the greatest effect on the separation. [2,70] Adding miscible counter ions such as ammonium formate or ammonium acetate have been shown to improve peak shapes.…”

“…[26,34,[46][47][48][49][50][51][52][53][54][55] Its wide applicability and demonstrated analytical and preparative benefits have helped define SFC as the new high speed paradigm in analytical chemistry. While the topic of high throughput (HT) SFC analysis and purification has been covered in several additional reviews, [24,52,[56][57][58][59][60][61] the focus of this review will discuss the factors to consider before contemplating a HT SFC application, advances in stationary phases, and those emerging applications that have the potential to expand the use of SFC into the future.…”

Design and Synthesis of Protein Superfamily-Targeted Chemical Libraries for Lead Identification and Optimization

“…Certainly, the questionable analytical purity of many of the originally-generated libraries in the field -particularly those produced in the order of tens or hundreds of thousands as part of mixture synthesis, a ubiquitous though unreliable approach during the earlier years -has since, not surprisingly, come under intense scrutiny. However, the advent of purification techniques, such as solid phase extraction [21], high-throughput preparative HPLC [22], and supercritical fluid chromatography [23], has enabled the rapid, parallel synthesis of smaller, though chemically-pure libraries of the type now routinely produced within most research physiological and pathophysiological roles [33][34][35][36]. The mechanism of action of GPCRs is well established: agonistinduced activation of GPCRs causes a conformational change within the cytoplasmic domain which subsequently results in the interaction of the receptor with guanine nucleotidebinding regulatory ('G-') proteins, leading to a cascade of intracellular signal transduction events.…”