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Micro-fluidic chip

A microfluidic chip and pipeline technology, applied in the field of biomedical testing, can solve problems such as product blanks, reduce the amount of reagents, and improve the detection range.

Pending Publication Date: 2022-05-17
FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, the detection of CAR-T cells and cytokines in China is mainly through flow cytometry and traditional ELIS

Method used

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Examples

Experimental program
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Example Embodiment

[0035] Example 1

[0036] The present embodiment provides a microfluidic chip (length× width× height of 10cm×8cm×1cm), for simultaneous detection of CAR-T cells and a variety of cytokines in the patient's peripheral blood, including: from top to bottom sequentially connected valve layer, pipe layer and basal layer;

[0037] Wherein, the pipe layer as Figure 1-2 Shown, comprising: sequentially connected sample mixing module 9, CAR-T cell capture detection module 6, concentration gradient generation module 8, cell filtration module 13 and cytokine detection module 15;

[0038] The sample mixing module 9 comprises: a sample mixing pipe in an "S" shape; the sample mixing pipe is provided with a first inlet for injecting dilution solution 1, a second inlet for injecting fluorescent antibody solution 2 and a third inlet for injecting a venous blood sample 3, the sample mixing pipeline is provided with a first outlet port 4;

[0039] The CAR-T cell capture detection module 6 compri...

Example Embodiment

[0050] Example 2

[0051] The present embodiment provides a method for simultaneously detecting CAR-T cells and a variety of cytokines in the peripheral blood of patients, the steps comprising:

[0052] One drop (50 μL) of peripheral blood sample, 50 μL of fluorescent antibody solution, and 50 μL of PBS dilution solution were injected at a flow rate of 5 μL / s from the third inlet 3, the second inlet 2 and the first inlet of the sample mixing module 9 as described in Example 1, respectively; in the sample mixing pipeline, the peripheral blood sample was diluted, and the CAR-T cells therein efficiently bound to the fluorescent antibody in the fluorescent antibody solution;

[0053]3-fold diluted peripheral blood sample in CAR-T cell capture detection module 6, T cells and CAR-T cells are captured by T cell capture antibodies, and then fluorescent CAR-T cells and T cells can be detected at the same time, and the proportion of CAR-T cells can be calculated;

[0054] Most of the T c...

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Abstract

The invention relates to a micro-fluidic chip, which is used for simultaneously detecting CAR-T cells and various cytokines in peripheral blood of a patient, and comprises a valve layer, a pipeline layer and a substrate layer which are sequentially connected from top to bottom, the pipeline layer comprises a sample mixing module, a CAR-T cell capturing and detecting module, a concentration gradient generating module, a cell filtering module and a cell factor detecting module which are sequentially connected through pipelines; the micro-fluidic chip can be used for simultaneously detecting CAR-T cells and cell factors in peripheral blood of a cancer patient receiving CAR-T cell therapy, a whole blood sample does not need to be transferred into a flow cytometer for detection, samples with different concentrations can be simultaneously detected in the micro-fluidic chip, the detection range is effectively expanded, and the detection cost is reduced. A reaction pipeline of the micro-fluidic chip is a closed environment, so that a relatively stable detection environment is provided for detection of CAR-T cells and cell factors; and the blood sample size required by detection is 50 microliters and is less than that required by the existing detection technology, and the dosage of corresponding reagents is also reduced.

Description

Technical field [0001] The present invention relates to the field of biomedical detection technology, in particular to a microfluidic chip. Background [0002] Chimeric Antigen Receptor T-Cell Immunotherapy (Chimeric Antigen Receptor T-CellImmunootherapy), or CAR-T therapy, is a new type of precision-targeted therapy for the treatment of tumors, which is reshaping the treatment of patients with advanced cancer, especially those with limited treatment options, and is effective in the treatment of relapsed, refractory B-cell non-Hodgkin lymphoma, and may be extended to other solid and liquid tumors. A fast, accurate new tumor immunotherapy, which is considered to have the potential to cure cancer, has brought new hope to cancer treatment. However, CAR-T therapy is also accompanied by a series of life-threatening adverse reactions such as cytokine release storms (CRS) and neurotoxicity (NE). The study found that the aggravation of NE is related to the high car-T cell replication le...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12M1/00B01L3/00
CPCC12M23/16C12M41/36C12M41/38B01L3/5027B01L2200/10
Inventor 荆雯雯程训佳冯萌张天然
Owner FUDAN UNIV
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