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Yeast surface display method based on digital microfluidic system

A yeast surface display and digital micro technology, which is applied in the field of biomedical detection and analysis, can solve the problems of long screening cycle and cumbersome steps of the yeast surface display system, and achieve the effects of shortening the operation cycle, saving training costs, and improving the process

Pending Publication Date: 2022-02-01
FOSHAN ACXEL BOXIN TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a yeast surface display method based on a digital microfluidic system. The technical problem to be solved is that the existing yeast surface display system has cumbersome steps and a long screening period.

Method used

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  • Yeast surface display method based on digital microfluidic system
  • Yeast surface display method based on digital microfluidic system
  • Yeast surface display method based on digital microfluidic system

Examples

Experimental program
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Effect test

Embodiment 1

[0054] Figure 1-21 The shown yeast surface display method based on the digital microfluidic system includes the following steps:

[0055] (1) Rabbits were immunized with IFN-γ, and the immunized serum, bone marrow and spleen were obtained, RNA was extracted and reverse transcribed, and the sequence of the antibody library was amplified by specific primers, and a yeast display library was constructed. Incubate the yeast strain in the yeast display library with biotinylated IFN-γ protein and streptavidin magnetic beads 11 to obtain a yeast library containing the complex of library yeast-biotinylated protein-magnetic beads 11 In liquid 1, the cells that can be recognized by the corresponding protein can form positive yeast complex 2, and the remaining negative yeast 3 cells that cannot be recognized by the corresponding protein will continue to maintain a free state.

[0056] (2) inject the liquid in step (1) into such as Figure 33 In the chip of the digital microfluidic syst...

Embodiment 2

[0064] A yeast surface display method based on a digital microfluidic system, comprising the following steps:

[0065] (1) Rabbits were immunized with IFN-γ, and the immunized serum, bone marrow and spleen were obtained, RNA was extracted and reverse transcribed, and the sequence of the antibody library was amplified by specific primers, and a yeast display library was constructed. Incubate the yeast strain in the yeast display library with biotinylated IFN-γ protein and streptavidin magnetic beads 11 to obtain a yeast library containing the complex of library yeast-biotinylated protein-magnetic beads 11 In liquid 1, cells that can be recognized by the corresponding protein can form positive yeast complex 2, and the rest of the negative yeast 3 cells that cannot be recognized by the corresponding protein will continue to maintain a free state;

[0066] (2) Inject the liquid in step (1) into the chip of the digital microfluidic system, turn on the electrode under the voltage of...

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Abstract

The invention provides a yeast surface display method based on a digital microfluidic system. The method comprises the following steps: integrating a magnetic adsorption module into a digital microfluidic chip system, combining yeast library bacteria with magnetic beads to form a compound, enriching yeast cells through a magnetic adsorption effect between a magnetic adsorption system and the magnetic beads, and then completing yeast surface display through labeling of a fluorescent substance labeled antibody and fluorescence detection. According to the yeast surface display method based on the digital microfluidic system, enrichment and screening processes of yeast surface display are integrated on the digital microfluidic chip, and a flow cytometer is not needed, so that costs for purchasing instruments, training personnel and the like are saved to a great extent; the whole process is highly integrated, high in automation degree and relatively simple to operate; and the operation period is greatly shortened, and the overall time efficiency is improved.

Description

technical field [0001] The invention belongs to the field of biomedical detection and analysis, and in particular relates to a yeast surface display method based on a digital microfluidic system. Background technique [0002] Yeast surface display is a protein eukaryotic surface display technology. First, connect the exogenous gene to a specific surface display vector, transform yeast cells, and induce the expression of the exogenous target protein. The expressed fusion target protein is fixed on the yeast cell wall by interacting with specific proteins on the yeast cell wall, so as to achieve the purpose of displaying foreign protein molecules on the yeast cell surface. Yeast display technology can be applied to many aspects at present, providing new solutions for protein evolution, antibody screening, antibody affinity, etc. [0003] The traditional yeast display technology needs to construct a protein display library, and induce the yeast to display the protein on the s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02C12M1/00G01N21/64G01N33/53
CPCC12N1/02C12M23/16C12M33/00G01N33/53G01N21/6486
Inventor 不公告发明人
Owner FOSHAN ACXEL BOXIN TECH CO LTD
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