title: Supercritical fluid assisted production of micrometric powders of the labile trypsin and chitosan/trypsin composite microparticles.
abstract: Supercritical fluid assisted atomization introduced by a hydrodynamic cavitation mixer (SAA-HCM) was used to prepare micrometric particles of a labile protein, i.e., trypsin from aqueous solution without use of any organic solvents. The trypsin particles precipitated had various morphologies under different process conditions, with particle diameters ranging from 0.2 to 4 μm. FTIR, SDS-PAGE, CD and fluorescence spectra were performed to analyze the structural stability of the protein, and trypsin retained above 70% of the biological activity. Besides, chitosan was selected as the polymer carrier in an effort to prepare trypsin composite microparticles via SAA-HCM process. The influences of chitosan molecular weight, polymer/protein ratio and solution concentration on the particle morphology and size distribution were investigated in detail. Non-coalescing spherical composite microparticles with a narrow particle distribution (0.2-3 μm) could be obtained. The SAA-HCM prepared particles were amorphous as demonstrated by XRD and had a loading efficiency about 90%. The protein release profiles of the composite microparticles were evaluated using both the immersion condition and a Franz diffusion cell. Finally, the distribution of the protein within the particles was characterized through CLSM analysis of FITC-labeled trypsin-loaded chitosan microparticles. The SAA-HCM process is demonstrated to be a protein-friendly and promising technique for production of protein and polymer/protein composite particles formulations from aqueous solutions for drug delivery systems.
authors.affiliation: College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Yu-Bin
authors.initials: YB
authors.lastname: Shen
authors.affiliation: College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Yi-Xin
authors.initials: YX
authors.lastname: Guan
authors.affiliation: College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China. Electronic address: yaosj@zju.edu.cn.
authors.forename: Shan-Jing
authors.initials: SJ
authors.lastname: Yao
mesh_terms: D002245:Carbon Dioxide / Q000737:chemistry; D048271:Chitosan / Q000737:chemistry*; D002942:Circular Dichroism; D003692:Delayed-Action Preparations / Q000737:chemistry; D065546:Drug Liberation; D004591:Electrophoresis, Polyacrylamide Gel; D010316:Particle Size; D011208:Powders; D017550:Spectroscopy, Fourier Transform Infrared; D013818:Thermogravimetry; D014357:Trypsin / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: D003692:Delayed-Action Preparations; D011208:Powders; D002245:Carbon Dioxide; D048271:Chitosan; D014357:Trypsin
keywords: Activity; Chitosan; Composite microparticles; Hydrodynamic cavitation mixer; Supercritical fluid assisted atomization; Trypsin