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Multi-chromaticity-electrophoresis separating and detecting system based on array capillary electrofocasing

An electrophoretic separation and detection system technology, which is applied in the direction of material separation, measurement devices, and analysis materials, can solve problems such as the impact of multi-dimensional system resolution, and achieve the effect of improving separation flux

Inactive Publication Date: 2005-01-05
FUDAN UNIV
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Problems solved by technology

In this way, the resolution of the next dimension cannot be very high due to the speed limit, and the resolution of the entire multi-dimensional system will be affected accordingly.

Method used

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  • Multi-chromaticity-electrophoresis separating and detecting system based on array capillary electrofocasing
  • Multi-chromaticity-electrophoresis separating and detecting system based on array capillary electrofocasing

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Experimental program
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Embodiment Construction

[0025] 1. RPLC generally uses a 250 μm capillary with a flow rate of 2 μl / min. Determine the number and interval of intercepted fractions according to the sample condition and gradient time.

[0026] 2. Dozens of (generally 40-100) quartz capillaries with a length of 20.0 cm and an inner diameter of 75 μm remove 4-8 cm (for example, 6 cm) of the outer wall coating in the central part as a detection window, and engrave on both ends of the detection window Scratches, but doesn't break. Fast-drying glue is used to fix the capillary on the two grooves of the insulating material substrate, and the quick-drying glue is applied around the groove. Then fix a plastic fence and platinum wire on the groove to facilitate the storage of electrolyte and high pressure.

[0027] 3. After the capillary is fixed, gently break it at each scratch, and cover the broken part with a layer of porous polymer film (such as cellulose acetate film).

[0028] 4. After the RPLC fraction is mixed with th...

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Abstract

The invention is a multi-dimension chromatogram, electrophoresis separation and detecting system based on array capillary equipotential focus. The system integrates the reverse light phase chromatogram and capillary pipe equipotential focus, the first dimension is the capillary pipe reverse chromatogram carrying high efficient separation to the complex protein mixture, and fraction separated flows into the second dimension-capillary pipe equipotential focus to be separated with higher efficiency and high flux quickly. Compared with single-channel multi-dimension chromatogram system, the design of the array is more proper to the analysis of complex sample with large scale, and the peak capacity of the two-dimension system can be upgraded further. It is excellent at aspect of sample condensing and low rich degree protein detection, and the CIEF can be connected with the mass spectrum online to carry on sample identification.

Description

technical field [0001] The invention is an efficient and fast multi-dimensional chromatography-electrophoresis separation and detection system for complex protein samples. technical background [0002] Multidimensional chromatography / electrophoresis separation is a new type of proteomics research method developed in recent years. It combines the chromatographic modes with two orthogonal separation mechanisms. After the previous one-dimensional separation, all components enter the next dimension, and the samples that are not completely separated in the previous one-dimensional are further separated. Its resolving power is much higher than that of single-dimensional chromatography or electrophoresis. [0003] Reversed-phase liquid chromatography (RPLC) and capillary isoelectric focusing (CIEF) are two high-resolution modes for the separation of biological samples with orthogonal separation mechanisms. RPLC is the most commonly used separation mode, which can withstand high i...

Claims

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Application Information

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IPC IPC(8): G01N30/38
Inventor 张祥民毛煜
Owner FUDAN UNIV
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