Method for detecting monoclonal antibody and application thereof

A monoclonal antibody and antibody technology, applied in the field of cell analysis, to reduce the reaction process, accurate and reliable identification results, and reduce interference.

Active Publication Date: 2020-12-15
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above technologies still rely on fluorescent signals to discover specific, high-affinity, functional antibodies from a large number of cell populations

Method used

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  • Method for detecting monoclonal antibody and application thereof
  • Method for detecting monoclonal antibody and application thereof
  • Method for detecting monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] This embodiment provides a method for detecting monoclonal antibodies, the process is as follows figure 1 As shown, the specific steps include:

[0060] (1) Prepare cells

[0061] Training sample cells: Obtain living cells and / or known cell lines, and prepare single-cell suspensions with buffer. The buffer is 0.85% NaCl physiological saline, phosphate buffer or other buffer solutions suitable for the physiological concentration of the cells. Two processing methods: 1. Add one or more detection antigens and incubate at 4°C, 37°C or room temperature for 1 hour; 2. Prepare single-cell suspension directly without adding antigens. Then wash twice with buffer, and resuspend cells with cell culture medium to prepare a single cell suspension for later use. The cell culture solution is RPMI 1640, DMEM or other nutrient solutions suitable for the growth of the cells.

[0062] Detection of sample cells: Obtain blood, tissues and organs of immunized mice, rabbits and other anim...

Embodiment 2

[0100] In this embodiment, using the method in Example 1, the hybridoma cells that secrete anti-CD45 antibody and the Sp2 / 0 cells that do not secrete antibodies are taken as examples, and the specific application methods are described as follows:

[0101] All the experimental steps are carried out in the cell room in a sterile environment, the consumables and instruments are sterilized in advance, and the reagents are also sterile.

[0102] Collect the hybridoma cells and Sp2 / 0 cells, centrifuge at 1000rpm for 3min, resuspend the cells with RPMI1640 culture medium, adjust the cell concentration to 10 per ml 6 cells. 100ul of cell suspensions were taken respectively, added to 96-well plates, and rested for 10 minutes. Spectral information of cells is captured with hyperspectral imaging under a microscope. 10 images were collected for each type of cell. Each type of cell is divided into training set, verification set and inspection set. In the training set, the cells that sec...

Embodiment 3

[0105] In this embodiment, the method of Example 1 is used to screen the cells that secrete PDL1 for the lymphocytes of mice, and the specific application methods are described as follows:

[0106] All the experimental steps are carried out in the cell room in a sterile environment, the consumables and instruments are sterilized in advance, and the reagents are also sterile.

[0107] Take two mice, one is a normal mouse, and the other is a mouse immunized with programmed death protein ligand 1 (PD-L1) protein. The spleen cells of the mice were extracted, ground aseptically, washed twice with serum-free medium, and centrifuged at 1000rpm for 3min to remove fat and other adherents. B cells were isolated from the spleen suspension using mouse B lymphocyte separation medium. Resuspend B cells in RPMI 1640 medium and adjust cells to 10 6 cells. 100ul of cell suspensions were taken respectively, added to 96-well plates, and left to stand for 10 minutes. Cell spectral information...

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Abstract

The invention relates to a method for detecting a monoclonal antibody and application thereof, and the method for detecting the monoclonal antibody comprises the following steps: acquiring hyperspectral data of a cell to be detected through hyperspectral imaging detection, and inputting the hyperspectral data into a machine learning model to obtain an antibody secretion type of the cell to be detected. By introducing a hyperspectral imaging technology and a microfluidic technology, single cell separation is realized, a hyperspectral image of a single cell is rapidly acquired, required featureinformation is rapidly and accurately mined from a large amount of hyperspectral data by using a machine learning algorithm, and high-precision identification and classification of single cells are automatically realized. The analysis process has the characteristics of high flux and intelligence, high result accuracy, and high sensitivity, and a rapid, non-contact and damage-free identification method is provided for antibody drug discovery and cell analysis.

Description

technical field [0001] The invention relates to the field of cell analysis, in particular to a method for detecting monoclonal antibodies and its application. Background technique [0002] Monoclonal antibody drugs have specific targeting, high sensitivity, and high affinity, and are often used in other clinical diseases such as tumors, autoimmunity, and infectious diseases, as well as in the field of diagnostic testing. In the research and development of antibody drugs, obtaining antibodies with high specificity, high affinity, strong competitiveness, and neutralizing activity is a key step and evaluation index in the development of antibody drugs. [0003] At present, the methods for preparing antibodies include hybridoma technology, single B cell technology, antibody library (phage library, ribosome library, artificially synthesized antibody library, etc.) B lymphocytes (bone marrow, peripheral blood, lymph nodes, spleen and other tissues and organs) are isolated from mi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/25G01N21/75G16B20/50G16B40/00
CPCG01N21/25G01N21/75G16B40/00G16B20/50
Inventor 陶然张卫凯李勤吕蒙李伟魏泽文
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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