Kit and extraction method for high-flux rapid magnetic-bead-method extraction of proteome

Abstract

The invention discloses a kit and extraction method for high-flux rapid magnetic-bead-method extraction of a proteome. The kit comprises the following reagents and materials: a protein reducer, a protein alkylation reagent, magnetic beads capable of adsorbing proteins and peptide segments, proteolytic enzyme, a porous plate and a porous magnetic frame matched with the porous plate. The extraction method disclosed by the invention comprises the steps of reductive alkylation of protein liquid, magnetic-bead-added protein adsorption, twice zymolysis, peptide segment adsorption of magnetic beads, and peptide segment eluting. Compared with the prior art, the kit and the extraction method, disclosed by the invention, have the advantages that the problems in the traditional methods that peptide segment demineralization steps are tedious and are long in consumed time and high in consumable cost can be thoroughly solved, and high-flux proteome extraction can be achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit and an extraction method for extracting proteomes by a high-throughput rapid magnetic bead method. Background technique [0002] Protein is the material basis of life and the main bearer of life activities. Therefore, proteomics, which focuses on the change of protein in the whole organism, has attracted much attention. As a large-scale protein research method, proteomics is widely used in food, animal and plant, clinical research and other fields. As the main technical means of proteomics research, mass spectrometry needs to analyze the protein in the sample before detection. Perform pretreatment for instrument detection, the quality and speed of protein pretreatment directly affect the entire research process. [0003] General mass spectrometry proteome pretreatment steps include protein extraction, concentration detection, reductive alkylation, protein precipita...

AI Technical Summary

Problems solved by technology

The whole process is complicated and cumbersome. The pretreatment process requires a lot of manual operations on a single sample, and the centrifuge tube where each sample is located is constantly opened, closed and replaced. The number of samples processed by one experimenter in a single batch generally does not exceed 24; The desalted samples generally use solid phase extraction to remove impurities and desalt. Each sample needs a solid phase extraction cartridge or Tip (10-30 minutes / sample), and the desalted samples need to be concentrated and replaced. (3-10 hours) before mass spectrometry detection can be performed. It takes 3-5 days to process a batch of samples, which takes a long time; and due to many processing steps and large losses, a large initial sample volume is required. For clinical samples Or other precious samples, often because of insufficient sample size, the quality of the experiment is affected or even impossible

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