High-throughput protein

KeyWords Adenovirus reporter protein luciferase green fluorescent protein high-throughput cell-based screening. 

Another example is the purification of a P-lactam antibiotic, where process-scale reversed-phase separations began to be used around 1983 when suitable, high pressure process-scale equipment became available. A reversed-phase microparticulate (55—105 p.m particle size) C g siUca column, with a mobile phase of aqueous methanol having 0.1 Af ammonium phosphate at pH 5.3, was able to fractionate out impurities not readily removed by hquid—hquid extraction (37). Optimization of the separation resulted in recovery of product at 93% purity and 95% yield. This type of separation differs markedly from protein purification in feed concentration ( i 50 200 g/L for cefonicid vs 1 to 10 g/L for protein), molecular weight of impurities (<5000 compared to 10,000—100,000 for proteins), and throughputs ( i l-2 mg/(g stationary phasemin) compared to 0.01—0.1 mg/(gmin) for proteins). 

Mac Beath G, Schreiber SL (2000) Printing proteins as microarrays for high throughput function determination. Science 289 1760-1763... 

Structural genomics aims to use high-throughput structure determination and computational analysis to provide structures and/or 3D-models of every tractable protein. The intention is to determine as many protein structures as possible and to exploit the solved structures for the assignment of biological function to hypothetical proteins. 

Vangrevelinghe E, Zimmermann K, Schoepfer J, Portmann R, Fabbro D, Furet P. Discovery of a potent and selective protein kinase CK2 inhibitor by high-throughput docking. J Med Chem 2003 46 2656-62. 

Doman TN, McGovern SL, Witherbee BJ, Kasten TP, Kurumbail R, Stallings WC, Connolly DT, Shoichet BK. Molecular docking and high throughput screening for novel inhibitors of protein tyrosine phosphatase IB. J Med Chem 2002 45 2213-21. 

Wilkins MR et al High-throughput mass spectrometric discovery of protein post-translational modifications. J Mol Biol 1999 289 645. 

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