Detection method and application of human peripheral blood mononuclear cell membrane surface transporter

Abstract

The invention relates to the technical field of cell membrane protein detection, in particular to a detection method and application of a human peripheral blood mononuclear cell membrane surface transporter. The method comprises the following steps: (1) separating human peripheral blood mononuclear cells; (2) extracting peripheral blood mononuclear cell membrane protein; (3) performing membrane protein pretreatment; and (4) performing nanoliter liquid phase tandem high-resolution mass spectrometry detection, and calculating the expression quantity of the transporter on the surface of the PBMC membrane. The method not only can be used for detecting the transporter on the surface of the PBMC membrane of the organ transplantation patient, but also can be used for detecting the transporter on the surface of the PBMC membrane of other immune disease patients; and the transporter on the surface of the PBMC membrane can be detected only by 1-3 mL of a whole blood sample, multiple transporters can be detected at the same time, and the flux is high.

Description

technical field [0001] The invention relates to the technical field of cell membrane protein detection, in particular to a detection method and application for detecting transporters on the membrane surface of human peripheral blood mononuclear cells. Background technique [0002] After organ transplantation, immunosuppressants need to be used to prevent the occurrence of organ rejection. Immunosuppressants used for organ transplantation mainly include glucocorticoids, calcineurin inhibitors, anti-proliferation and anti-metabolic inhibitors, and antibody-based biological agents . Immunosuppressants enter lymphocytes and bind to tacrolimus binding protein (FKBP12), mTOR and other proteins in lymphocytes to exert pharmacological effects, and the formed complexes such as TAC-FKBP12 and Sirolimus-FKBP-mTOR inhibit the participation of T-cells Activated calcineurin, inhibits T-cell activation and thus exerts immunosuppressive effects. The concentration of immunosuppressant in l...

AI Technical Summary

Problems solved by technology

[0005] Traditional methods for detecting membrane proteins include ELISA, Western Blotting, flow cytometry and other technologies. Traditio

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