Antibody biological activity detection method based on surface plasma resonance technology

Abstract

The invention provides an antibody biological activity detection method based on a surface plasma resonance technology, and particularly relates to the technical field of biological detection, human Burkitt's lymphoma cell strain Daudi cells highly expressed by an LAG3 receptor are selected to be combined with a C1 chip, and the combined cells are fixed on the surface of the chip; when the LAG3 iscombined with cells on the surface of the chip, a response signal is captured through a biomacromolecule interaction instrument Biacore T200; when the anti-LAG3 antibody is added, the combination ofLAG3 and cells is reduced, the response value is reduced, and the antibody activity at the cellular level is realized by comparing the response value so as to realize rapid detection. The method is simple in step, easy to operate, capable of achieving real-time dynamic monitoring and high in efficiency, has a specific recognition effect, and has great advantages in antibody drug screening, batch release, stability monitoring, even process development and other applications of immune checkpoint targets.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to an antibody biological activity detection method based on surface plasmon resonance technology. Background technique [0002] Immune checkpoint blocking antibodies can fight cancer by enhancing the function of the immune system, and this type of cancer immunotherapy has become one of the important methods in the treatment of various malignant tumors in recent years. Immune checkpoints can be stimulatory or inhibitory, with co-stimulatory proteins signaling to promote an immune response to pathogens, and inhibitory in the opposite direction. Among them, the inhibitory molecules cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and programmed cell death receptor 1 (PD-1) are the two most studied immune checkpoint molecules. CTLA-4 is the first immune checkpoint target for clinical use, and its inhibitor ipilimumab was approved for marketing in 2011. Pr...

AI Technical Summary

Problems solved by technology

However, none of the existing detection methods can meet the needs of rapid detection of in vitro activity of antibodies targeting immune checkpoints, for example, mixed lymphocyte reaction (Mixed lymphocyte reaction, MLR) and antigen-specific stimulation assays (antigen-specific recall assays ) can detect biological activity by detecting the secretion of IFN-γ, but the detection cycle is longer and takes 4-5 days

Moreover, these methods use primary cells. Due to the differences brought about by different individuals and the complexity of experimental operations, the variability of these two detection methods is very large.

In addition, in vitro activity detection methods based on T cell proliferation, cytotoxicity, etc. also require the use of primary cells and complicated operations, which are not satisfactory as a rapid detection method for the biological activity of immune checkpoint antibodies.

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