Method for detecting cell PD-L1 protein expression in body fluid sample and special reagent thereof

A PD-L1, body fluid sample technology, applied in the field of protein immunohistochemical detection, can solve the problems that have not been mentioned yet, and achieve the effect of simple operation and high sensitivity

Inactive Publication Date: 2019-07-05
NINGBO MEIJING MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the immunohistochemical method (used to detect PD-L1 antibody) is suitable for tissues, and it has not been mentioned whether it can be used for the detection of PD-L1 protein expression in tumor cells such as pleural effusion, bone marrow, and peripheral blood

Method used

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  • Method for detecting cell PD-L1 protein expression in body fluid sample and special reagent thereof
  • Method for detecting cell PD-L1 protein expression in body fluid sample and special reagent thereof
  • Method for detecting cell PD-L1 protein expression in body fluid sample and special reagent thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Taking the peripheral blood samples of patients with lung adenocarcinoma as an example, using the CellRich immunomagnetic particle capture instrument of Ningbo Meijing Medical Technology Co., Ltd. TM and its supporting reagents to capture tumor cells. And use the detection method of the present invention to carry out follow-up detection to it, concrete steps are as follows:

[0029]1. Preparation of immunomagnetic particles (take 2mg magnetic particles as an example)

[0030] 1) Pretreatment of magnetic particles: select brown round (or oval) magnetic particles with a particle size of 200nm, and a carboxyl content of 200-400μmol / g; add 1mL 0.05mol / L MES to a centrifuge tube containing 2mg of magnetic particles Buffer solution (acidic buffer solution), let stand on the magnetic stand for 3 minutes, aspirate and discard the supernatant, add 1mL 0.05mol / L MES buffer solution to the centrifuge tube, after ultrasonic treatment, let stand on the magnetic stand for 3 minutes,...

Embodiment 2

[0045] Taking the ascites sample of a patient with lung adenocarcinoma as an example, the detection method of the present invention detects it.

[0046] 1. Preparation of immunomagnetic particles (take 2mg magnetic particles as an example)

[0047] 1) Pretreatment of magnetic particles: select brown round (or oval) magnetic particles with a particle size of 200nm, and a carboxyl content of 200-400μmol / g; add 1mL 0.05mol / L MES to a centrifuge tube containing 2mg of magnetic particles Buffer solution (acidic buffer solution), let stand on the magnetic stand for 3 minutes, aspirate and discard the supernatant, add 1mL 0.05mol / L MES buffer solution to the centrifuge tube, after ultrasonic treatment, let stand on the magnetic stand for 3 minutes, Discard the supernatant and repeat 2-4 times in total; add 0.5mL 0.05mol / LMES buffer into the centrifuge tube again, and continue to sonicate in the ultrasonic instrument to obtain a magnetic particle suspension with consistent size, compl...

Embodiment 3

[0062] Capture and enrichment of leukemia cells in patients' bone marrow: Firstly, the patient's bone marrow samples were subjected to density gradient centrifugation to collect mononuclear cells in the samples, and the collected cells were resuspended with the pretreatment solution and then used the company's immunomagnetic particle capture instrument CellRich TM And its matching reagents are used to capture leukemia immature cells.

[0063] 1. Preparation of immunomagnetic particles (take 2mg magnetic particles as an example)

[0064] 1) Pretreatment of magnetic particles: The selected magnetic particles have a particle size of 1 μm, a carboxyl content of 200-600 μmol / g, a round shape, and a brown appearance; add 1 mL of 0.05 mol / L to a centrifuge tube containing 2 mg of magnetic particles MES buffer (acid buffer), let stand on the magnetic stand for 3 minutes, aspirate and discard the supernatant, add 1mL 0.05mol / L MES buffer to the centrifuge tube, after ultrasonic treatme...

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Abstract

The invention discloses a method for detecting a cell PD-L1 protein expression in a body fluid sample and a special reagent thereof. The method comprises the steps that a prepared immunomagnetic particle mixture and tumor circulating cells in the body fluid sample are combined and placed in an immune magnetic particle trap for capturing; and the captured tumor cells are stained by a PD-L1 immunohistochemical staining reagent to evaluate the PD-L1 protein expression, wherein the immunomagnetic particle mixture is coated with at least one of anti-human EpCAM immunomagnetic particles, anti-humanICAM-1 immunomagnetic particles, anti-human EGFR antibody immunomagnetic particles, anti-human folate receptor immunomagnetic particles, anti-human CD10 immunomagnetic particles, anti-human CD19 immunomagnetic particles, anti-human CD20 immunomagnetic particles and anti-human CD33 immunomagnetic particles. The method has the advantages of simple operation, high sensitivity, and easy detection result interpretation.

Description

technical field [0001] The invention belongs to the field of immunohistochemical detection of proteins, and in particular relates to a method for detecting cell PD-L1 protein expression in body fluid samples and special reagents thereof. Background technique [0002] PD-1 is a type I transmembrane immunoglobulin receptor, a member of the CD28-mediated immune response superfamily, and an important inhibitory molecule on the surface of T cells. Its intracellular segment contains an ITIM and an immune receptor tyrosine conversion Motif (ITSM), ITSM mediates the recruitment of protein tyrosine phosphatase family phosphatases and the inhibition of T cell activation signals. The receptors of PD-1 include PD-L1 (B7-H1) and PD-L2 (B7-DC), which mainly play an important role in the tumor microenvironment in the effector phase of the immune system. PD-L1 is inducibly expressed in a variety of tumor cells and hematopoietic cells in the tumor microenvironment. By combining with PD-1, i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/54326G01N33/6854
Inventor 张晓晶沈挺宋毅
Owner NINGBO MEIJING MEDICAL TECH
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