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Biomarker related to tumor immunotherapy effect and application of biomarker

A biomarker and immunotherapy technology, applied in the application field of predicting the effect of tumor immunotherapy, can solve problems such as adverse reactions, single biomarkers, test result sampling time and sampling site effects, etc.

Active Publication Date: 2020-06-26
HANGZHOU KAIBAOLUO BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the use of PD-L1 expression as a predictor of ICI efficacy has the following defects: a considerable proportion of patients with advanced tumors cannot provide enough tumor tissue for the detection of PD-L1 expression; There will be heterogeneity in the region, and the detection results may be affected by the sampling time and sampling location; in addition, the differences in detection methods, evaluation standards, and the definition of positive cut-off values ​​will also lead to different judgments of results, which all lead to PD. - L1 expression does not serve as a sufficiently comprehensive independent biomarker for clinical treatment decisions
However, it is worth noting that so far, in terms of the relationship between the expression of autoantibodies against tumor-associated antigens and ICI response, the relevant reports are still very limited; moreover, these autoantibodies are all single biomarkers, predicting It may not be accurate, and its actual prediction effect still needs further verification
[0009] Immune checkpoint inhibitors are very expensive drugs, unlike conventional chemotherapy, although immune checkpoint inhibitor drugs are effective for some patients, they can also cause serious adverse reactions, especially the development of systemic autoimmune diseases

Method used

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  • Biomarker related to tumor immunotherapy effect and application of biomarker
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  • Biomarker related to tumor immunotherapy effect and application of biomarker

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Experimental program
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Effect test

preparation example Construction

[0159] (1) Preparation and immobilization of antigenic protein

[0160] The cDNA of tumor-associated antigen (TAA) was cloned into PET28(a) expression vector containing 6XHis tag. At the N-terminal or C-terminal of the antigen, streptavidin or the like (biotin-binding tag protein) is introduced. The obtained recombinant expression vector was transformed into Escherichia coli for expression. After the protein was expressed in the inclusion body, the protein was denatured with 6M guanidine hydrochloride, and refolded in vitro according to the standard method, and then Ni-NTA affinity was carried out through the 6XHis tag Column purification to obtain antigenic protein.

[0161] (2) Preparation of plasma

[0162] Venous blood was drawn in EDTA-treated or citric acid-treated blood collection tubes within one week to one day before immunotherapy. Then centrifuge at 1000-2000RCF at room temperature for 15 minutes; after centrifugation, gently transfer the supernatant to another c...

Embodiment 1

[0199] In order to find autoantibodies that can indicate the effect of immunotherapy, firstly, the plasma of 47 normal healthy people (healthy control group) and 47 patients diagnosed with lung cancer (lung cancer group) was used to detect whether there were autoantibodies against the purified antigenic protein in lung cancer patients . The healthy control group consisted of those who were not diagnosed with cancer in the past and present. The lung cancer group consisted of patients diagnosed with lung cancer, including 10 patients with small cell lung cancer, 12 patients with lung squamous cell carcinoma, 19 patients with lung adenocarcinoma, and 6 patients with other subtypes of lung cancer. Plasma was drawn before treatment. The experimental population information is shown in Table 1.

[0200] Table 1. Primary screening test crowd information

[0201]

[0202]

[0203] In order to compare the concentrations of autoantibodies corresponding to each antigen in the heal...

Embodiment 2

[0208] Autoantibodies are detected in the baseline (before treatment) plasma of immunotherapy patients (Table 3), and the antigenic proteins of the first three groups (sensitivity>5%) in Table 2 of Example 1 are used to screen for autoantibodies related to the curative effect of lung cancer patients. .

[0209] In order to screen immunotherapy predictive markers suitable for clinical application, this study included various scenarios of immunotherapy for lung cancer patients as much as possible. There are 38 lung cancer patients who use immunotherapy as first-line treatment, while 40 lung cancer patients have undergone one or more treatments such as chemotherapy and targeted therapy, and then choose immunotherapy (post-line treatment). In addition, the immunotherapy used by the enrolled patients included immune checkpoint inhibitors, including imported nivolumab, pembrolizumab, and domestic immune checkpoint inhibitors (Table 3). Whether it is first-line or later-line immunot...

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Abstract

The invention relates to a biomarker related to a tumor immunotherapy effect and an application thereof, and discloses a biomarker for predicting the tumor immunotherapy effect, and the biomarker is an autoantibody of a group of tumor-associated antigens. The biomarker can be detected in a sample from a tumor patient to predict the clinical effect of administration of immunotherapy, thereby facilitating the determining of whether the tumor patient can benefit from immunotherapy. The invention also provides an antigen protein combination for detecting the biomarker, a kit containing the antigenprotein combination, and a corresponding detection or diagnosis method.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular, the present invention relates to a biomarker related to the effect of tumor immunotherapy, a protein combination for detecting it and corresponding application in predicting the effect of tumor immunotherapy. Background technique [0002] Immunotherapy is currently the most promising research direction in the field of tumor treatment. One of the hotspots is the use of immune checkpoint inhibitors (immune checkpoint inhibitor, ICI) immune blockade therapy, such as based on blocking programmed death factor-1 (programmed death factor-1) Death-1, PD-1) / programmed death ligand-1 (programmed deathligand-1, PD-L1) immune checkpoint pathway therapy. [0003] The blocking therapy of the PD-1 / PD-L1 immune checkpoint pathway usually refers to the injection of specific antibodies against PD-1 or PD-L1 into tumor patients, so that the tumor no longer has the ability to escape the attack of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57488G01N33/57423
Inventor 孙苏彭杨盼盼隗啸南周静康美华孙立平
Owner HANGZHOU KAIBAOLUO BIOLOGICAL SCI & TECH
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