Preparation method of micro-fluidic chip interface for two-dimensional protein electrophoretic separation

Abstract

The invention discloses a preparation method of a micro-fluidic chip interface for two-dimensional protein electrophoretic separation, and relates to a micro-fluidic chip. The method comprises the following steps of: preparing polyacrylamide gel in a vertical channel of the micro-fluidic chip and filling a buffering aqueous solution; introducing an organic phase through a horizontal channel of the micro-fluidic chip and filling the horizontal channel; introducing an organic phase into which tetraisopropyl titanate is dissolved and filling the horizontal channel; draining an organic solution of tetraisopropyl titanate out of the horizontal channel, and cleaning the horizontal channel; adding an isoelectric focusing buffer solution into which proteins and amphoteric electrolytes are dissolved into the horizontal channel, applying electric fields to the two ends of the horizontal channel for performing isoelectric focusing, and realizing first-dimension separation of proteins in the horizontal channel according to respective different isoelectric points; and after finishing isoelectric focusing, applying electric fields to the two ends of the horizontal channel for performing second-dimension gel electrophoresis, making micropores on a TiO2 film pass through an electric field, and making the proteins enter a second-dimension channel by passing through the micropores on the TiO2 film under the action of the electric field for performing two-dimension separation.

Description

technical field [0001] The invention relates to a microfluidic chip, in particular to a method for preparing a microfluidic chip interface used for protein two-dimensional electrophoresis separation. Background technique [0002] Protein is the most important carrier and functional executor of life activities. Proteomics (Proteome), first proposed in 1994, refers to all proteins that can be expressed by genes, that is, the expression of cells or tissues or organisms at a specific time and space. All protein. Exploring the mode of action, functional mechanism, regulation and regulation of proteins, and the interactions within protein groups can provide a theoretical basis and basis for clinical diagnosis, pathological research, drug screening, new drug development, and metabolic pathways. The main methods for proteomics research include two-dimensional electrophoretic separation, mass spectrometry, microarray chips, etc. (Domon, B., Aebersold, R., Science 2006, 312, 212.; Ku...

AI Technical Summary

Problems solved by technology

However, there are still the following shortcomings: time-consuming and labor-intensive, poor repeatability, and insufficient separation ability for extreme proteins, low-abundance proteins, and hydrophobic proteins. These shortcomings seriously hinder the further rapid development of related research.

However, these methods still have certain shortcomings: the operation is cumbersome and complicated, the pore size is too large, and the resistance is too small, which cannot simply and effectively reduce the spontaneous diffusion between two-dimensional channels.

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