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An immunosensor system for the detection of PD-L1 for non-disease diagnostics

A PD-L1, immunosensor technology, applied in instruments, measuring devices, scientific instruments, etc., can solve problems such as affecting the enhancement effect of SERS, and achieve the effect of reducing steric hindrance interference, realizing directional combination, and increasing the capture amount.

Active Publication Date: 2022-06-28
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Factors such as the material of the SERS active substrate, the shape and size of the nanoparticles, the adsorption amount and the distance of the probe on the active substrate will all affect the enhancement effect of SERS.

Method used

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  • An immunosensor system for the detection of PD-L1 for non-disease diagnostics
  • An immunosensor system for the detection of PD-L1 for non-disease diagnostics
  • An immunosensor system for the detection of PD-L1 for non-disease diagnostics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Preparation of Homing Peptide-functionalized magnetic-coated silver nanoparticles, namely Ag@MNPs-HomingPeptide:

[0045] The synthesis steps of Homing Peptide-functionalized magnetic-coated silver nanoparticles are as follows: after washing about 1 mL of magnetic-coated silver nanoparticles solution (Ag@MNPs) with PBS for three times, add specific 10 μL 1 mg / mL Homing Peptide, the magnetic-coated silver nanoparticles The volume ratio with the specific Homing Peptide is about 2:1, and the reaction time is about 4 hours. After that, the magnetic separation is performed, the supernatant is discarded, and resuspended in PBS to obtain Homing Peptide-functionalized magnetic-coated silver nanoparticles (Ag@ MNPs-HomingPeptide).

[0046] The obtained Ag@MNPs-Homing Peptide, which can be abbreviated as Ag, was characterized by ultraviolet-visible spectroscopy (Uv-vis), Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), Zeta potential and t...

Embodiment 2

[0047] Example 2 Preparation of an immunosensor system for detecting PD-L1

[0048] First, mix 100 μL of 2 mM diamine guest with 400 μL of pSC 4 -AuNPs reacted for 10 minutes to form corresponding 3D-AuNPs as Figure 4 shown, and added to the attached pSC 4 on the chip surface as Figure 5 The 3D-AuNPs showed the sensitizing effect of SPR signal and electrochemical signal. in a multiparameter surface plasmon resonance instrument (MP-SPRModelNavi TM 210A), 500 μL of 1 μg / mL PD-L1 antibody was added to react with 3D-AuNPs at a flow rate of 2 μL / min, and then the surface of the gold sheet was washed with buffer to elute the unconnected antibody. Then, the chip was reacted with 1 mg / mL BSA solution for half an hour to avoid nonspecific adsorption on the chip surface. Meanwhile, Ag@MNPs-Homing Peptide was reacted with different concentrations of analytes at 25 °C for 30 min, and then separated by external magnetism. Finally, the post-reaction Ag@MNPs-Homing Peptide was added ...

Embodiment 3

[0049] Example 3 Monitoring PD-L1 using the immune sensing system SPR

[0050] The concentrations of PD-L1 used were: 1ng / ml, 10ng / ml, 100ng / ml, 200ng / ml, and 300ng / ml.

[0051] Test conditions: Using an SPR instrument, running at the same flow rate, SPR measurements at different concentrations of PD-L1 were performed at room temperature.

[0052] like Image 6 As shown in A, as the concentration of PD-L1 increases, the resulting change in the SPR angle is also greater.

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Abstract

The invention discloses an immunosensor system for detecting PD-L1 for non-disease diagnosis, which includes diamine guest molecules, pSC 4 ‑AuNPs, pSC attached 4 chip, PD‑L1 antibody, and Homing Peptide functionalized magnetic-coated silver nanoparticles, in which diamine guest molecules interact with pSC 4 ‑AuNPs attached with pSC 4 The chip surface self-assembled to construct 3D-AuNPs as a sensitizing layer, PD-L1 antibody was adsorbed on the sensitizing layer, and Homing Peptide functionalized magnetic-coated silver nanoparticles were added to the attached pSC after reacting with the analyte. 4 The surface of the chip can be specifically recognized with the PD‑L1 antibody. The 3D‑AuNPs of the present invention are used as the antibody immobilization substrate, and the supramolecular layer can directionally immobilize the antibody. Based on the mechanism of the intracellular peptide, it can replace the antibody to build an immune sensor, and the cost is low.

Description

technical field [0001] The invention relates to the field of immunotherapy, in particular to an immunosensor system for detecting PD-L1 for non-disease diagnosis. Background technique [0002] The immune cells of a healthy body can find and kill cancer cells, but under the induction of various innate and acquired factors, the immune system will lose its killing ability, leading to the occurrence and development of cancer. "Cancer immunotherapy" is a treatment method that uses the body's own immune system to attack cancer cells. [0003] Programmed death ligand 1 (PD-L1) is one of the major molecules regulating T cell immune responses. PD-L1 (B7-H1) belongs to the same CD28 / B7 / CTLA-4 family member as CD80 and CD86, and the coding gene is located on the ninth chromosome p24. It is a type I membrane protein consisting of a single hydrophobic transmembrane domain, a short intracellular domain, and two Ig-like domains within the extracellular domain, an N-terminal IgV domain an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/553G01N33/543G01N27/26G01N21/552
Inventor 陈红霞
Owner SHANGHAI UNIV