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Tumor tissue in-vitro pathological staining kit

A staining reagent and tumor tissue technology, applied in the field of biomedicine, can solve the problems of limited indications, high price, and unclear side effects of targeted drugs.

Inactive Publication Date: 2017-06-13
INST OF AUTOMATION CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In recent years, with the rapid development of tumor genomics and molecular pharmacology, the research and development of new tumor-targeted drugs has achieved good research results, but many targeted drugs have limited indications, are expensive, and have not very side effects. clear

Method used

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  • Tumor tissue in-vitro pathological staining kit
  • Tumor tissue in-vitro pathological staining kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Targeting and specificity detection of Cetuximab

[0046] 1. Preparation of orifice plate to be tested

[0047] 1. Cell plating

[0048] a) Prepare the complete medium required for each cell.

[0049] Specifically, the basic medium RPMI or DMEM accounts for 80%-90%, fetal bovine serum accounts for 10%-20%, and double antibody stock solution (penicillin+streptomycin) is added at a volume fraction of 1%, and penicillin and streptomycin are used. The final concentrations of the proteins were 100 U / mL and 100 μg / mL, respectively.

[0050] b) Before the experiment starts, confirm the name of the cell line marked on the 100mm culture dish, the culture medium, passage time, generation and other information to ensure that the experiment is correct.

[0051] c) Aspirate the cell culture medium with a sterile pipette.

[0052] d) Rinse the cell surface with 3-5 mL of sterile PBS solution, and then use a sterile pipette to suck out the PBS waste.

[0053] e) Add 1 m...

Embodiment 2

[0087] Example 2 Cetuximab is used for pathological staining of tumor tissue in vitro

[0088] 1. Preparation of the test drug Cetuximab-FITC

[0089] Preparation of Cetuximab-FITC

[0090] Cetuximab was dispersed into PBS with a pH of 8.5, and FITC was added in a molar ratio of 1:1 to 1:3, and reacted at 35° C. for 2 hours. Cetuximab-FITC complex was separated and purified by PD10 desalting column. Its specific concentration and connection efficiency were analyzed by HPLC.

[0091] 2. The specific implementation steps of in vitro pathological staining of tumor tissue are as follows:

[0092] 1) The tissue to be tested is removed from the animal or human body and soaked in 10% neutral formalin solution for 1-2 days;

[0093] 2) Embed the tissue in paraffin, cut it into thin slices with a thickness of 5 μm with a pathological microtome, dewax the slices, and wash with distilled water;

[0094] 3) Stain with 0.001-0.005 mg / mL Cetuximab-FITC staining solution at room temperatu...

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Abstract

The invention discloses a tumor tissue in-vitro pathological staining kit. The tumor tissue in-vitro pathological staining kit is a kit developed aiming at leading Cetuximab to be used in new applications of tumor tissue in-vitro pathological staining, experiments verification finds that after the Cetuximab is subjected to fluorescent labeling and then used for tumor tissue in-vitro pathological staining, normal tissues and tumor tissues can be well distinguished, well-differentiated and poorly-differentiated tumor tissues can be distinguished to certain extent, and the kit can be used for tumor detection, thus realizing new use for approved drugs.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a tumor tissue in vitro pathological staining kit. Background technique [0002] At present, tumor has become a global disease that threatens human health. The development of efficient detection drugs for early tumor, micrometastasis and lymph node metastasis is very important to prolong the survival period and quality of life of patients. [0003] In recent years, with the rapid development of tumor genomics and molecular pharmacology, the research and development of new tumor-targeted drugs has achieved good research results, but many targeted drugs have limited indications, are expensive, and have not very side effects. clear. Cetuximab, known as "cetuximab" in Chinese, belongs to chimeric IgG1 monoclonal antibody, and its molecular target is epidermal growth factor receptor (EGFR). Drugs mainly used to treat advanced colon cancer. The anti-tumor mechanism of the drug i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30
Inventor 田捷靳玉慎马喜波孟慧张帅
Owner INST OF AUTOMATION CHINESE ACAD OF SCI
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