Mobility electrophoresis based method for measuring size of material

A substance and size technology, applied in the field of determination of substance size based on mobility electrophoresis, which can solve the problems of complicated processing steps and large amount of reconstructed data.

Pending Publication Date: 2018-10-16
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When using the above techniques to analyze the structure, it is necessary to obtain high-purity and large-scale samples first, the pre-p

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  • Mobility electrophoresis based method for measuring size of material
  • Mobility electrophoresis based method for measuring size of material

Examples

Experimental program
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Effect test

Embodiment 1

[0088] The structure of somatostatin was determined by mobility electrophoresis.

[0089] Mobility electrophoresis experiment conditions: the sample is 1mg / mL somatostatin, the buffer is 50% aqueous methanol (w:w contains 0.1% formic acid, buffer pH 3.0), and 0.5% (w:w) phenol is added as neutral Marker, mix well. The length of the capillary tube is 50cm, the distance from the injection end to the detection window is 40cm, the inner diameter of the tube is 75μm, and the wavelength of 214nm is detected by ultraviolet spectrophotometry. The operation method is as follows: use 1000mbar air pressure to feed the buffer solution for 180s to flush the inner pipe of the capillary; use 50mbar air pressure to inject the sample for 5s; apply an air pressure of 60mbar at both ends of the capillary, and the separation voltage is -20kV. Experimental results such as Figure 4 (a) shown. According to the sample equivalent spherical radius characteristic curve (formula 4) under the experime...

Embodiment 2

[0092] The structure of angiotensin I was determined by mobility electrophoresis.

[0093] Mobility electrophoresis experiment conditions: the sample is 1 mg / mL angiotensin Ⅰ, the buffer is 50% methanol aqueous solution (w:w contains 0.1% formic acid, buffer pH 3.0), and 0.5% (w:w) phenol is added as the medium Sex markers, mix well. The length of the capillary tube is 50 cm (the distance from the injection end to the detection window is 40 cm), the inner diameter of the tube is 75 μm, and the wavelength of 214 nm is detected by ultraviolet spectrophotometry. The operation method is: use 1000mbar air pressure to inject buffer solution for 180s to flush the separation channel; use 50mbar air pressure to inject samples for 5s; apply air pressure at both ends of the separation channel to 60mbar, and the separation voltage is -20kV. Repeat this operation 5 times, the experimental results are as follows Figure 6 As shown in (c), the repeatability of 5 groups of parallel experiment...

Embodiment 3

[0096] The structure of bradykinin was determined by mobility electrophoresis.

[0097] Mobility electrophoresis experimental conditions: the sample is 1mg / mL bradykinin, the buffer is 50% aqueous methanol (w:w contains 0.1% formic acid, buffer pH 3.0), and 0.5% (w:w) phenol is added as neutral Marker, mix well. The length of the capillary tube is 50 cm (the distance from the injection end to the detection window is 40 cm), the inner diameter of the tube is 75 μm, and the wavelength of 214 nm is detected by ultraviolet spectrophotometry. The operation method is: use 1000mbar air pressure to inject buffer solution for 180s to flush the separation channel; use 50mbar air pressure to inject samples for 5s; apply air pressure at both ends of the separation channel to 60mbar, and the separation voltage is -20kV. Repeat this operation 5 times, the experimental results are as follows Figure 7 As shown in (c), the repeatability of 5 groups of parallel experiments is good. Calculat...

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Abstract

The invention relates to the technical field of mobility electrophoresis and discloses a mobility electrophoresis based method for measuring the size of a material. The method comprises using an injection pump to inject a buffer solution into a sampling end of a capillary tube, maintaining for a first period of time, then using a sampling pump to inject a sample solution into the sampling end of the capillary tube, using the injection pump to inject the buffer solution into the sampling end of the capillary tube again after finishing sampling, applying a separation voltage between the two endsof the capillary tube at the same time; using a detector to detect a detection window of the capillary tube, acquiring the appearance time tM of a neutral marker and the appearance time t of a sample; acquiring a characteristic curve of the equivalent sphere radius of the sample, further acquiring the ionic equivalent sphere radius R of the sample; acquiring the ellipse reconstruction limit curveof the sample according to the ionic equivalent sphere radius R of the sample, and further acquiring the conformation distribution of the sample under the experiment condition. The operation of the method is convenient, the analysis speed is fast, and the analysis can be conducted together with the separation of the mixture.

Description

technical field [0001] The invention relates to the technical field of mobility electrophoresis, in particular to a method for determining the size of a substance based on mobility electrophoresis. Background technique [0002] Biological macromolecules are closely related to life activities, and the analysis of the structure of macromolecules is helpful to understand their functions and mechanisms. Common structural elucidation techniques include X-ray crystallography, nuclear magnetic resonance spectroscopy, and cryo-electron microscopy. When using the above-mentioned techniques to analyze the structure, it is necessary to obtain high-purity and large-scale samples first, the pre-processing steps are relatively cumbersome, and the reconstruction after data collection also faces problems such as a large amount of data. Therefore, there is a need for a solution with high sensitivity and the ability to analyze the structure of complex components. In addition, the use of auxi...

Claims

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Application Information

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IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 徐伟贺木易张荣楷
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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