Solution-based screening

Micro reactors are continuous-flow devices consuming small reaction volumes and allowing defined setting of reaction parameters and fast changes. Hence they are ideal tools for process screening and optimization studies to develop solution-based chemistries. 

Sundberg, S.A. High-throughput and ultra-high-throughput screening solution- and cell-based approaches. Curr. Opin. Biotechnol. 2000, 11, 47-53. 

Flarakos, J., Morand, KL., Vouros, P. High-throughput solution-based medicinal library screening against human serum albumin. Anal. Chem. 2005, 77, 1345-1353. 

S.L. Childs, N. Rodriguez-Hornedo, L.S. Reddy, A. Jayasankar, C. Maheshwari, L. McCausland, R. Shipplett, B.C. Stahly, Screening strategies based on solubility and solution composition generate pharmaceutically acceptable cocrystals of carbamaze-pine, CrystEngComm 10 (2008) 856-864. 

Sundberg, S. A. High-Throughput and Ultra-High-Throughput Screening Solution- and Cell-Based Approaches. 

Before crystallization trials, the protein was subjected to gel filtration on Superdex-75 (Pharmacia) in 50 mM sodium/potassium phosphate buffer, pH 7.4, containing 1 mM EDTA, 50 mM 2-mercaptoethanol, 150 mM sodium chloride, 5% glycerol and 5% 2-propanol, as described previously (12). The statine-based inhibitor, LP-149 (Ac-Nal-Val-Sta-Glu-Nal-NH2 e Nal is naphtylalanine and Sta is statine) (Fig. 1), was prepared at Lilly Research Laboratories (K. Hui, unpublished results). Crystallization was carried out at 4 °C using the hanging-drop vapor diffusion method as follows 2.5 //I of the FIV PR(D30N) at 7 mg/ml complexed with LP-149 (1 4 molar ratio) in 50 mM imidazole-HCl pH 7.0 containing ImM EDTA and 1 mM dithiothreitol were mixed with an equal volume of 2 M ammonium sulfate, 0.1 M sodium acetate, pH 4.6 (Hampton Crystal Screen, solution 47). Crystals appeared within a few days and reached the size of 0.2 x 0.2 X 0.4 mm in one week. 

For the dioxin binder screening, some changes were made as shown in Fig. 8.6. A new pentapeptide library was prepared because a strong affinity is required, to detect dioxins at the ppb level. The design of the screening solution is very important since it determines which peptides can be screened. Also, the composition of the screening solution may restrict the detection conditions of the sensor. For the herbicide binder screening, ethanol was used to dissolve the herbicide in an aqueous buffer. The refractive index of ethanol (1.3623) is not very different from that of water (1.3335) therefore, the ethanol-based buffer solution could be used in the... 

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