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Detection material, detector and detection method for circulating tumor cells

A technology for detecting materials and tumor cells, applied in the field of detecting materials for circulating tumor cells, can solve the problems of strong non-specific adhesion, poor detection effect, weak adhesion, etc. mild conditions

Active Publication Date: 2022-05-13
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the existence of tumor heterogeneity, the adhesion of liver cancer CTCs on the bioorthogonal gold surface is weak and the non-specific adhesion is strong. The detection of liver cancer by gold surface has poor detection effect and high false positive

Method used

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  • Detection material, detector and detection method for circulating tumor cells
  • Detection material, detector and detection method for circulating tumor cells
  • Detection material, detector and detection method for circulating tumor cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] This embodiment provides a detection material for circulating tumor cells, which is a bio-orthogonal / anti-adhesion bifunctional chitosan coating. Wherein, "bioorthogonal" refers to groups that can undergo bioorthogonal reactions, including eight-membered ring octynes ​​and ortho-substituted triphenylphosphine groups used in Staudinger-Bertozzi ligation. "Anti-adhesion" refers to a functionalized chitosan coating having both a hydrophilic chitosan structure and a hydrophobic bioorthogonal structure. The preparation method of bio-orthogonal / anti-adhesion bifunctional chitosan coating comprises the preparation of chitosan coating and the functional modification of chitosan coating, specifically as follows:

[0067] Preparation of chitosan coating: Dissolve 95% deacetylated chitosan in 90% acetic acid solution; 4000 rpm spin coating on the substrate with hydrophilic surface to obtain chitosan coating, or use A chitosan coating was prepared on a substrate with a hydrophilic...

Embodiment 2

[0076] This embodiment provides a detection material for circulating tumor cells, which is a bio-orthogonal / anti-adhesion / reversible release trifunctional chitosan coating. Wherein, "bioorthogonal" refers to groups that can undergo bioorthogonal reactions, including eight-membered ring octynes ​​and ortho-substituted triphenylphosphine groups used in Staudinger-Bertozzi ligation. "Anti-adhesion" refers to a functionalized chitosan coating having both a hydrophilic chitosan structure and a hydrophobic bioorthogonal structure. "Reversible release" refers to having a reversible cleavage group, thereby releasing captured circulating tumor cells. The preparation method of bio-orthogonal / anti-adhesion / reversible release three-functional chitosan coating includes preparation of chitosan coating and functional modification of chitosan coating, specifically as follows:

[0077] Preparation of chitosan coating: Dissolve 95% deacetylated chitosan in 90% acetic acid solution; 2.4000 rpm ...

Embodiment 3

[0086] This example provides the whole process of liver cancer CTC detection based on bio-orthogonal / anti-adhesion dual-functional chitosan coating. step by step ( figure 1 A). Specific steps are as follows:

[0087] Clinical blood sample pretreatment: Collect 5ml of patient blood from an anticoagulant tube, use density gradient centrifugation to initially enrich CTCs, and use red blood cell lysate to remove residual red blood cells.

[0088] Manual labeling for metabolic glycoengineering: use 100uM Ac 4 ManAz is subjected to 48-hour metabolic sugar engineering treatment, and the treatment method refers to Chinese patent CN113358865A.

[0089] Co-incubation of the detector and the sample: In 1ml of DMEM medium, the metabolic glycoengineering sample was mixed with about 200mm 2 The detector is co-incubated (still or slowly flowed through), and other blood components that do not adhere to the detector are washed away to obtain CTCs that adhere to the surface of the detector....

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Abstract

The invention discloses a circulating tumor cell detection material, a detector and a detection method, the circulating tumor cell detection material is fixed on a substrate, and the detection material is chitosan modified by a functional group; the functional group comprises a biological orthogonal reaction group; or comprises a bio-orthogonal reactive group and a reversible reactive group. The circulating tumor cell detector comprises a substrate and the detection material fixed on the substrate. According to different modification degrees, the novel material for detecting the circulating tumor cells can achieve biological orthogonal / anti-adhesion dual functions or biological orthogonal / anti-adhesion / reversible release three functions, chitosan is used for replacing gold to serve as a functional surface, and the cost is reduced; the surface of the biological orthogonal chitosan has a good anti-adhesion effect, so that the detection accuracy is improved, and the probability of occurrence of false positive detection results is reduced.

Description

technical field [0001] The invention belongs to the technical field of biological materials, and in particular relates to a detection material, a detector and a detection method for circulating tumor cells. Background technique [0002] In 1869, Australian doctor Ashworth first discovered and proposed the concept of circulating tumor cells (Circulating Tumor Cell, CTC) in the blood of breast cancer patients. In 1976, Nowell revised the definition of CTC as: tumor cells originating from the primary tumor or metastatic tumor, gaining the ability to escape from the basement membrane and invade blood vessels through the tissue matrix. At present, CTC refers to the general designation of various tumor cells existing in peripheral blood, and its growth and dissemination run through all stages of tumor growth. [0003] Liver cancer is one of the six most common tumors and ranks among the top three in terms of mortality among all cancers. Early diagnosis and treatment are the diff...

Claims

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Application Information

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IPC IPC(8): C08B37/08G01N33/543G01N33/569G01N33/574
CPCC08B37/003G01N33/54393G01N33/56966G01N33/57438
Inventor 王怀雨劳智奇李伟
Owner SHENZHEN INST OF ADVANCED TECH