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Kit for guiding medication of tumor targeted drug

A technology of tumor targeting and kits, which is applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of differences in treatment effects, etc., and achieve the effects of less denaturation and inactivation, good stability, and enhanced probe sensitivity

Active Publication Date: 2016-06-15
SHANGHAI MAJORBIO BIO PHARM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to individual differences in tumor cells among different patients, there are also differences in the expression of target proteins in tumor cells, which makes the therapeutic effect of the same targeted drug different for different patients.

Method used

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  • Kit for guiding medication of tumor targeted drug
  • Kit for guiding medication of tumor targeted drug
  • Kit for guiding medication of tumor targeted drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1H

[0041] Embodiment 1 HER2-oligonucleotide probe

[0042] (1) Aldehyde modification of oligonucleotide probes

[0043] 150 nmol of Oligo1 (oligonucleotide) was prepared into a solution with 0.1 M pH 7.4 phosphate buffer. Weigh 1500nmol of SFB (N-succinimidyl 4-formylbenzoate), dissolve it with anhydrous DMF (N,N-dimethylformamide), react at room temperature for 2.5h, and purify through a column to obtain Oligo -FB (aldehyde modified oligonucleotide).

[0044]Detection of the concentration of Oligo-FB: detection of A by Nanodrop spectrophotometer 260 The calculated value of Oligo-FB is 2.6nmol / μL.

[0045] Detection of aldehyde group modification rate: Use quantitative 2-hydrazinopyridine-2-hydrochloride solution to detect the aldehyde group modification rate. Take the above-mentioned Oligo-FB and add it to the 2-hydrazinopyridine-2-hydrochloride solution, shake and mix well, and react at 37°C for 1 hour. The absorbance value at 360nm detected by Nanodrop is 6.3, and the modi...

Embodiment 2

[0058] The QPCR amplification specific detection of embodiment 2Oligo molecule

[0059] The Oligo1-13 molecules were diluted to three concentrations of 25000 molecules / μL, 83333 molecules / μL and 250000 molecules / μL. Then, taking Oligo1-3 as an example, Oligo1-3 was mixed at the same concentration to obtain mixed samples A, B, and C, as shown in Table 3.

[0060] Table 3 Example 2 sample formula

[0061]

[0062]

[0063] The above samples were configured with QPCR amplification solution according to Table 4 to obtain a QPCR amplification kit, and then Oligo1-3 was subjected to QPCR amplification according to the procedures shown in Table 5, and Oligo1-3 in mixed samples A, B, and C Each was subjected to QPCR amplification, using the extended Oligo1-3 as a template, and the Ct values ​​of the amplification results are shown in Table 6. Among them, the extension primer is RT-P, its 3' end is complementary to the 3' end of the oligonucleotide, the 5' end can form a hairpi...

Embodiment 3

[0078] Embodiment 3 makes standard curve

[0079] The same result can be obtained with Oligo1-13. In order to simplify the process, the following examples use Oligo1-4 as an example.

[0080] According to the steps of Example 1, Oligo2 was modified with SFB having a molar equivalent of 5 times to obtain Oligo-FB, and SANH with a molar equivalent of 10 times was used to modify EGFR with a hydrazine group to obtain EGFR-SANH, and the molar ratio was 10:1 EGFR-Oligo2 was obtained after reacting Oligo-FB with EGFR-SANH at room temperature for 16 hours.

[0081] According to the steps of Example 1, Oligo3 was modified with 20-fold molar equivalent of SFB to obtain Oligo-FB, and PD-L1 was hydrazino-modified with 50-fold molar equivalent of SANH to obtain PD-L1-SANH. The molar ratio PD-L1-Oligo3 was obtained after 8:1 Oligo-FB and PD-L1-SANH were reacted at room temperature for 24 hours.

[0082] According to the steps of Example 1, Oligo4 was modified with 10-fold molar equivalent...

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Abstract

The invention provides a kit for guiding the medication of a tumor targeted drug. The kit comprises an antibody-oligonucleotides probe obtained by coupling an antibody corresponding to a target and oligonucleotides, an amplification primer for performing PCR amplification to the antibody-oligonucleotides and a buffer solution, and a fluorescent probe. According to the kit, the antibody corresponding to the target on tumor cells is coupled with oligonucleotides so as to be used as the probe, and the expression of the target antigen can be detected by virtue of PCR, so that a more appropriate target can be selected to guide the medication of the targeted drug.

Description

technical field [0001] The invention relates to the field of multiple immunoassays, in particular to a kit for guiding the administration of tumor targeting drugs. Background technique [0002] In the past 20 years, targeted drugs have gradually become the mainstay of cancer treatment. Experiments in vitro and in animals have shown that these targeted drugs can induce apoptosis of cells, and through complement-mediated cytotoxicity (complement-mediated cytotoxicity, CMC) and antibody-dependent cell-mediated cytotoxicity (antibody -dependent cellular cytotoxicity, ADCC) kills target cells. Compared with traditional chemotherapy drugs, targeted drugs can kill tumor cells with high selectivity and reduce the damage to normal tissues. They have the characteristics of low toxicity and high efficiency, and may fundamentally inhibit or eliminate tumor cells. [0003] Generally speaking, there are usually multiple available targets on the surface of a tumor cell, and there are dif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57488
Inventor 邓文斌李静陈昌岳蔡红东甘广利张祥林
Owner SHANGHAI MAJORBIO BIO PHARM TECH
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