High Throughput Crystallization of Peptides Using the iCrystal System and Metal-Assisted and Microwave-Accelerated Evaporative Crystallization (MA-MAEC) Technique

摘要

Metal-Assisted and Microwave-Accelerated Evaporative Crystallization (MA-MAEC) is a novel technique in the field of evaporative crystallization led by the Aslan Research Group. The MA-MAEC technique is based on combined use of microwave heating (speeds up the process) and metal nanoparticle structure such as silver, gold, copper, Indium tin oxide (ITO), nickel (provides selective nucleation sites) to yield quality crystals of individual amino acids, small molecules, and peptides in a very short period of time.;This Master of Science thesis focuses on the crystallization of glutathione (GSH), a tripeptide on 95-well iCrystal plates using a mono-mode microwave cavity (the iCrystal system) via MA-MAEC technique to yield high quality crystals. The previous studies of GSH crystallization using the MA-MAEC technique with Silver Nano films (SNFs)-deposited 21-well iCrystal platform proved most effective based on the crystallization time and the overall quality of GSH crystals at 500 mg/mL concentration. Sodium acetate was chosen to be the best solvent for crystallization. In this study, we put together the underlying mechanism studied in the prior research of GSH crystallization to successfully perform the crystallization on an ITO plated 95-well iCrystal plate at 500 mg/mL concentration at 5, 10, 30, 60 and 120 minutes time intervals hereby to enhance the crystallization process. Time of crystallization, crystal size, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) were used to characterize crystals.;The results demonstrated that the MA-MAEC technique affords for the rapid crystallization of GSH in a high-throughput fashion using the iCrystal mono-mode microwave system. The crystallization time was reduced by up to 3-fold compared to control conditions (room temperature). Crystal morphology of the product crystals was similar to those reported in the literature.