| Background: When using genetic engineering method to produce Human recombinant hemoglobin (human rHb) from Escherichia coli. There occurs a partly of abnormal heme inserted recombinant hemoglobin, In order to do investigation in raise up the proportion of normal recombinant hemoglobin and decrease the abnormal recombinant hemoglobin proportion in this technology. Firstly, we need to have a good method for their proportion analysis.Methods: In this paper, it will study on: (1) non-denaturing gel electrophoresis and (2) capillary zone electrophoresis (CZE). And the preparation of sample is used Escherichia coli containing plasmid PHE7 to produce the human recombinant hemoglobin. After that, do purification and two chromatographic steps are employed. The first column captures a large amount of the unwanted bacterial proteins while the hemoglobin passes through. Although this column can be run at higher pH to capture the rHb, nucleic acids and most bacterial proteins are considerably more acidic than rHb and tend to displace it off the resin. The second column further removes bacterial proteins. Alternatively, the SP column will not employ in here. And the sample after the second column will give to do the proportional analysis method development directly. 4%, pH 6.8 stacking gel and 12%, pH 7.66 resolving gel without SDS is involved in the non-denaturing gel electrophoresis. The proteins are stacked at pH 6.8 and resolved at pH 7.66. Running at a constant current of 100 mA. An uncoated silica capillary of 50 cm (40 cm to the detector) × 75 μm is involved in capillary zone electrophoresis (CZE). The buffers have used were: 0.1M Tris-HCl, in pH 8.00, pH 8.25, pH 8.50, pH 8.75, and pH 9.00. Peaks detect at 415 nm. The sample injects under 0.5psi for 7s and electrophoreses at 15kV for about 25 minutes.Results: There is any separation can obvious in the result of non-denaturing gel electrophoresis under this condition. Oppositely, it shows a trend to separate in pH 9.0...
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