Detecting method for circulating tumor cell surface marker molecule PD-L1

Abstract

The invention discloses a detecting method for a circulating tumor cell surface marker molecule PD-L1. The detecting method comprises the following steps: (1) treating whole blood with red blood celllysate to separate out karyocyte, and fixing with formaldehyde; (2) performing positive screening through tumor immune fluorescent marker cytokeratin antibody anti-CK, performing PD-L1 primary antibody incubation on all cells, and performing PD-L1 secondary antibody incubation with a fluorescence group marked with FITC, and marking all cells through cell nucleus fluorescent dye DAPI; and (3) adopting high-flux multi-color imaging analysis to select light filter plates of CY5, FITC and DAPI, observing fluorescent color on the surface of a channel, and finally realizing detection on the circulating tumor cell surface marker molecule PD-L1. The detecting method is simple and convenient, is quick, is economical, is non-invasive, is high in sensitivity and is good in specificity.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a method for detecting the surface marker molecule PD-L1 of circulating tumor cells. Background technique [0002] Programmed death receptor-1 (PD-1) is the main immune checkpoint receptor, by binding to its ligand programmed death ligand-1 (PD-L1), it down-regulates the effector function of T cells, thus contributing to To maintain tolerance to tumor cells. Blocking these pathways by anti-PD-1 and anti-PD-L1 antibodies may help prevent this downregulation and allow T cells to maintain their antitumor properties and ability to mediate tumor cell death. There are currently three main inhibitors of PD-1 and PD-L1 on the market, pembrolizumab (trade name: Keytruda), Nibolumab (trade name: Opdivo) and Atezolizumab (trade name: Tecentriq), which can be used for melanoma, non-small cell Treatment of various cancers such as lung cancer and bladder cancer. However, not all p...

AI Technical Summary

Problems solved by technology

At present, the detection of PD-L1 in my country is relatively chaotic. First, the staining techniques and conditions are not uniform; second, the staining antibodies are diverse; third, the pathological evaluation standards are not yet unified. The value of the level is reduced

In addition, because immunohistochemistry must obtain tissue for detection, whether it is through surgery or puncture, it is an invasive operation that will cause harm to the patient

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