Full-automatic protein purification system device and application thereof
A protein purification and system device technology, which is applied in the field of automatic protein purification system device, separation and purification, and achieves the effect of low quality requirements and increased scope of application
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Embodiment 1
[0038] Embodiment 1 automatic protein purification system device
[0039] figure 1 The fully automated protein purification system setup is shown. The fully automatic protein purification system device includes a chromatography column 0, a peristaltic pump 1, a silicone hose 2, a positioning column 3, a stepper motor 4, a first container 5, a second container 6, a third container 7, and a fourth container 8. Fifth container 9, sixth container 10, first two-way solenoid valve 51, second two-way solenoid valve 61, third two-way solenoid valve 71, fourth two-way solenoid valve 81, fifth two-way solenoid valve 91. The sixth two-way solenoid valve 101, the liquid level detector 11 and its circuit board control system, wherein the stepping motor 4 drives the positioning column 3 connected thereto, so that the positioning column can rotate 360 degrees; the silicone hose 2 One end is connected to the outlet of the chromatography column 0, the middle is stuck on the peristaltic pum...
Embodiment 2
[0042] Embodiment 2 control parameter design
[0043] There are three main operating parameters that can be set for the device of the present invention: the number of column hanging times N, the delay time T seconds and the liquid flow rate. N represents the number of times the sample liquid flows through the chromatography column, which is actually the number of adsorption between the sample and the medium. In the manual purification of protein chromatography, it is usually sufficient to hang the column twice for adsorption, so N can be directly set equal to 2. The delay time T seconds means that when the equipment is running, when the electrode on the A / B metal probe of the liquid level detector is just exposed to the liquid surface, the system will receive a signal from high level to low level, and it actually takes T seconds later, figure 1 The liquid level inside the middle chromatography column 0 can be lowered to the upper end position of the adsorption medium (indicat...
Embodiment 3
[0044] Example 3 Protein Affinity Adsorption
[0045] After the Escherichia coli cells with His*6 affinity purification tag protein were sonicated, the supernatant obtained by high-speed centrifugation at 16,000 g for 30 minutes was obtained to obtain a supernatant with a volume of 50 ml, and the supernatant was placed in figure 2 in the first container 5. The number of column hanging times N is set to 2, the flow rate is set to 1.5 ml / min, and the delay time T is equal to 30 seconds. The chromatographic column equipped with nickel medium was equilibrated with 50 mM Tris protein buffer (pH=8) in advance. After selecting the "adsorption" mode and starting the automatic program, the first two-way solenoid valve 51 connected to the first container 5 is powered on and connected to the pipeline, and the silicone hose 2 is positioned directly above the second container 6 . The peristaltic pump starts to drive the fresh protein liquid in the first container 5 to pass through the e...
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