Reassemblable quasi-chip free-flow electrophoresis with simple heating dispersion for rapid micropreparation of trypsin in crude porcine pancreatin.

Autor: Geng JZ; Laboratory of Analytical Biochemistry & Bio-separation, Shanghai Jiao Tong University, Shanghai, PR China., Shao J, Yang JH, Pang B, Cao CX, Fan LY
Jazyk: angličtina
Zdroj: Electrophoresis [Electrophoresis] 2011 Nov; Vol. 32 (22), pp. 3248-56.
DOI: 10.1002/elps.201100358
Abstrakt: 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.
(Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
Databáze: MEDLINE
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