title: Reassemblable quasi-chip free-flow electrophoresis with simple heating dispersion for rapid micropreparation of trypsin in crude porcine pancreatin.
abstract: An increasing number of small biosamples (e.g. proteins and enzymes) need micropreparation in lab. However, neither large-scale free-flow electrophoresis (LS-FFE) nor chip FFE (C-FFE) could fit the growing demands. Herein, a simple quasi-chip FFE (QC-FFE) was constructed. In contrast to C-FFE, the features of QC-FFE are as follows: (i) its separation chamber is reassemblable and rewashable avoiding discard of C-FFE due to blockage of solute precipitation in chamber; (ii) its chamber size is 45  mm × 30  mm × (80-500)  μm (108-654  μL volume) having function of micropreparation; (iii) there are up to 16 outlets in QC-FFE bestowing fine fraction for micropurification. The QC-FFE was used for the micropurification of model enzyme of self-digestible trypsin in crude pancreatin. Under the given conditions, the purification factor of enzyme was 11.7, the specific activity reached 6236  U/mg, the run time for 19  μL sample purification was 45  s and the throughput of trypsin was 3.34  mg/h, and the yield of pure trypsin was 55.2%. All of the results show the feasibility of enzyme micropreparation via QC-FFE. The developed device and procedure have potential use to other micropurification of protein or peptide sample.
authors.affiliation: Laboratory of Analytical Biochemistry & Bio-separation, Shanghai Jiao Tong University, Shanghai, PR China.
authors.forename: Jia-Zhen
authors.initials: JZ
authors.lastname: Geng
authors.forename: Jing
authors.initials: J
authors.lastname: Shao
authors.forename: Jing-Hua
authors.initials: JH
authors.lastname: Yang
authors.forename: Bo
authors.initials: B
authors.lastname: Pang
authors.forename: Cheng-Xi
authors.initials: CX
authors.lastname: Cao
authors.forename: Liu-Yin
authors.initials: LY
authors.lastname: Fan
mesh_terms: D000818:Animals; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004867:Equipment Design; D006863:Hydrogen-Ion Concentration; D010194:Pancreatin / Q000737:chemistry*; D013552:Swine; D014357:Trypsin / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D010194:Pancreatin; D014357:Trypsin