CN111624340A - Peripheral blood TCR marker of pancreatic cancer and detection kit and application thereof - Google Patents
Peripheral blood TCR marker of pancreatic cancer and detection kit and application thereof Download PDFInfo
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Abstract
The invention discloses a peripheral blood TCR marker of pancreatic cancer, a detection kit and application thereof. The marker comprises at least one protein with a sequence of SEQ ID NO. 1-100. Based on a high-throughput sequencing method, only a small amount of peripheral blood is needed to be taken, RNA is extracted, an immune map library is established through sample processing, and then high-throughput sequencing and TCR data analysis are performed, so that a characteristic TCR sequence in peripheral blood of pancreatic cancer is determined, and then a test result of a sample to be tested is compared with the characteristic TCR sequence, so that whether the pancreatic cancer is suffered or not is determined. The invention can simultaneously compare a great number of pancreatic cancer specific TCR sequences, has higher specificity and accuracy compared with the single detection of one or more markers, and improves the diagnosis efficiency.
Description
Technical Field
The invention belongs to the technical field of genetic engineering, and particularly relates to a peripheral blood TCR marker of pancreatic cancer, a detection kit and application thereof.
Background
Pancreatic cancer is a highly malignant, difficult to diagnose and treat malignant tumor of the digestive tract, and about 90% of ductal adenocarcinomas originate from the epithelial lining of the glandular duct. The incidence of pancreatic cancer varies from region to region, with an average of about 5.1/10 ten thousand people. The incidence is higher in men than in women, with a ratio of about 1.5-2: 1. The overall incidence of disease has been on the rise in recent years. The latest statistical data of the cancer center in China show that the pancreatic cancer is the 8 th of the incidence rate of the male malignant tumor in the Chinese city and the 5 th of the mortality rate of the malignant tumor in the people living in Beijing and Shanghai.
The etiology of pancreatic cancer is not well understood. Its occurrence is associated with smoking, drinking, high fat and high protein diets, excessive drinking of coffee, environmental pollution and genetic factors; recent survey reports show that the incidence rate of pancreatic cancer in the diabetic population is obviously higher than that of the common population; some people also pay attention to the fact that a certain relation exists between the chronic pancreatitis patient and the pancreatic cancer, and the pancreatic cancer occurrence proportion of the chronic pancreatitis patient is obviously increased; in addition, many factors are involved in the occurrence of the disease, such as occupation, environment, geography, etc. Common clinical manifestations of pancreatic cancer are as follows:
1. abdominal pain: pain is a major symptom of pancreatic cancer, whether the cancer is located in the head or tail of the pancreas. In addition to the pain in the middle or upper left and right abdomen, the chief complaints in a few cases are pain in the left and right lower abdomen, around the umbilicus or the whole abdominal pain, and even pain in the testis, which is easily confused with other diseases. When cancer involves the visceral envelope, peritoneum or retroperitoneal tissue, there may be tenderness at the corresponding site.
2. Jaundice: jaundice is an important symptom of pancreatic cancer, particularly pancreatic head cancer. Jaundice is obstructive with deep yellow urine and argillaceous stools, and is caused by the invasion or pressure of the lower end of common bile duct.
3. Digestive tract symptoms: the most common is loss of appetite, followed by nausea, vomiting, possibly diarrhea or constipation and even dark stools, with diarrhea often being steatorrhea.
4. Emaciation and lack of strength: pancreatic cancer is different from other cancers, and is often accompanied by emaciation and weakness in the early stage.
5. Abdomen mass wrapping: the deep pancreas is difficult to be touched in the back abdomen, and the abdominal mass is the result of the development of the cancer itself and is located in the lesion, such as the touched mass, which is usually in the advanced stage or late stage. The chronic pancreatitis can be seen as a mass, and is not easy to be distinguished from pancreatic cancer.
6. Symptomatic diabetes: the few patients initially develop diabetes, i.e., they develop diabetes before the onset of the main symptoms of pancreatic cancer, such as abdominal pain, jaundice, etc., so that the accompanying loss of lean mass and weight is mistaken for the manifestation of diabetes, regardless of pancreatic cancer.
7. Thrombophlebitis: patients with advanced pancreatic cancer develop migratory thrombophlebitis or arterial thrombosis.
8. Psychiatric symptoms: some patients with pancreatic cancer may exhibit psychiatric symptoms such as anxiety, impatience, depression, personality changes, and the like.
9. Ascites: generally, it is caused by peritoneal infiltration and diffusion of cancer in the late stage of pancreatic cancer.
10. And others: in addition, patients often complain of fever and marked weakness. It can be associated with high fever and even chill, which are similar to cholangitis, so it is easy to be confused with cholelithiasis and cholangitis.
Pancreatic cancer is clinically characterized by short course of disease, difficulty in early diagnosis, susceptibility to confusion among other diseases, and rapid development and deterioration of disease in middle and late stages. Therefore, treatment of pancreatic cancer is very difficult and prognosis is very poor. Because of the difficulty of early diagnosis, only 10% to 15% of patients have the opportunity for surgical resection. The average survival time of pancreatic cancer is 2-3 months after diagnosis, annual survival is 8%, and five-year survival rate is only 3%. Therefore, there is a great need to find effective diagnostic and therapeutic methods. At present, the diagnosis means aiming at pancreatic cancer is mainly divided into four categories of biochemical and molecular biological detection, gene detection, imaging detection and pathological detection.
1. Biochemical and molecular biological detection:
1) the carbohydrate antigen CA19-9 is the most common pancreatic cancer diagnostic marker at present, and has the following clinical characteristics: the sensitivity and specificity for diagnosing pancreatic cancer reach 78.2 percent and 82.8 percent respectively by taking the serum CA19-9>37U/ml as a positive index. About 10% of pancreatic cancer patients are Lewis Antigen negative, CA19-9 is not increased, and other tumor markers such as CA125 and/or CarcinoEmbryonic Antigen (CEA) are combined for auxiliary diagnosis. In the case of CA19-9 elevated patients, pancreatic cancer should be highly suspected after excluding factors such as biliary obstruction or biliary system infection.
2) Monitoring blood glucose changes: a) the elderly, those with low body mass index and new onset diabetes without family history of diabetes should be alerted to the onset of pancreatic cancer; b) patients suffering from diabetes in a long term, suffering from blood sugar fluctuation in a short term and difficult to control, should be vigilant of pancreatic cancer; c) the prospective research result shows that the incidence risk of pancreatic cancer is increased by 14% when the fasting blood sugar is increased by 0.56 mmol/L; 2) other biomarkers: microRNA, ctDNA, in-vitro Glypican-1 and the like in peripheral blood also have potential clinical application prospects, but clinical application is yet to be confirmed by high-level evidence-based medical evidence.
2. Gene detection: the gene mutations of CDKN2A, BRCA1/2, PALB2 and the like are proved to be closely related to the onset of familial pancreatic cancer. However, this gene test can only evaluate the risk of cancer and cannot be used as a diagnostic standard.
3. Imaging examination
1) Enhanced three-dimensional dynamic CT thin-layer scanning: at present, the most commonly used means for diagnosing pancreatic cancer can clearly display the size, position, density and blood supply condition of the tumor, and accordingly judge the adjacency relation between the tumor and blood vessels [ adopting CT (computed tomography Angiography, CTA) for inspection when necessary ] and adjacent organs, and guide the evaluation of the resectability of the tumor before operation and the effect of neoadjuvant chemotherapy.
2) Magnetic Resonance Imaging (MRI): besides displaying the anatomical characteristics of pancreatic tumors, the method can also clearly display whether metastatic lesions exist in the parapancreatic lymph nodes and the liver; and is superior to CT examination in differentiating edematous or chronic lump pancreatitis. The Magnetic Resonance pancreaticocholangiography (MRCP) and MRI thin-layer dynamic enhancement are combined for application, which is helpful for determining cystic and solid lesions of pancreas (particularly differential diagnosis of cystadenoma and papillary mucous tumor in pancreatic duct), further determining the expansion and invasion conditions of pancreatic duct and bile duct, and has higher diagnostic value.
3) Positron Emission Tomography (PET) -CT examination: can show the metabolic activity and metabolic load of the tumor, and has obvious advantages in the aspects of finding out the pancreatic extrapancreatic metastasis and evaluating the systemic tumor load.
4) The ultrasonic Endoscope (EUS) combines ultrasonic imaging on the basis of an endoscope technology, and improves the sensitivity and specificity of pancreatic cancer diagnosis; in particular, EUS guided Fine needle biopsy (EUS-FNA) has become the most accurate method for locating and qualitatively diagnosing pancreatic cancer. In addition, the EUS is also helpful for judging the tumor stage, the sensitivity and specificity for diagnosing pancreatic cancer at T1-2 stage are respectively 72% and 90%, and the sensitivity and specificity for diagnosing pancreatic cancer at T3-4 stage are respectively 90% and 72%.
4. And (3) pathological examination: histopathological and/or cytological examinations are "gold standards" for diagnosing pancreatic cancer. Except for patients who are planned to undergo surgical resection, the rest of patients should be strived for definitive pathological diagnosis before making treatment plans. Current methods for obtaining histopathological or cytological specimens include:
1) puncture biopsy under EUS or CT guidance; 2) examination of ascites abscission cytology; 3) probe biopsy under laparoscopic or open surgery.
However, the above conventional detection methods have their limitations. Such as biochemical and molecular biological detection, because the types of markers are few, it is difficult to ensure the specificity and sensitivity at the same time; imaging detection is expensive; the pathological examination needs puncture or ascites extraction, the patient has great pain, and the like. Moreover, all existing detection methods are difficult to distinguish early pancreatic cancer from other types of diseases, and patients often miss early diagnosis opportunities. Therefore, a pancreatic cancer diagnosis method with higher sensitivity and specificity and simple and easy operation is needed.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides the TCR marker of the peripheral blood of pancreatic cancer, the detection kit and the application thereof, and can accurately and quickly judge whether a patient with higher pancreatic cancer risk exists in a sample to be detected.
In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:
a peripheral blood TCR marker of pancreatic cancer comprises at least one protein shown as SEQ ID NO. 1-100 in sequence, and the specific sequence is shown in Table 1.
TABLE 1 marker sequences
Furthermore, the protein sequence of the marker is a protein with the same function expressed by the sequence shown in SEQ ID NO. 1-100 after one or more basic groups are substituted, deleted and/or replaced.
Further, the marker is a peripheral blood TCR CDR3 sequence.
The application of the marker in preparing a preparation for treating pancreatic cancer.
Further, the preparation includes a T cell receptor containing the marker, or a plasmid, viral vector or nucleic acid fragment capable of expressing the T cell receptor producing the marker.
A kit for pancreatic cancer detection comprising an antibody that specifically binds to the marker.
An agent comprising an antibody that specifically binds to the marker; the formulations can be used for diagnosis, prognosis, detection or screening of pancreatic cancer.
A protein chip for detecting pancreatic cancer comprises a substrate and a specific antibody spotted on the substrate, wherein the specific antibody is an antibody capable of specifically binding with the marker.
The principle of the invention is as follows: b-lymphocytes and T-lymphocytes in the human body are two important types of cells in the adaptive immune system. B is thinThe cell recognizes antigen through a cell surface B Cell Receptor (BCR), and later, the BCR expresses antibody and is secreted out of the cell when the B cell differentiates into plasma cells. T cells recognize antigens via T Cell Receptors (TCRs) on the cell surface. The diversity of BCRs and TCRs is the basis for establishing an adaptive immune system. The theoretical value of the diversity of BCR is 1018Theoretical value of TCR diversity is 1014. Among the BCR and TCR sequences, epitope 3(CDR3) is the most important part in determining the antigenic specificity, and therefore the sequence of CDR3 is considered to represent the properties of the BCR and TCR sequences.
In various diseases, the diversity or expression level of both BCR and TCR changes with different antigenic stimuli. Therefore, the occurrence and development of diseases can be tracked by using BCR or TCR high-throughput sequencing results. In human cells, after degradation of the senescent protein, fragments thereof are transported to the cell surface and presented to T cells in the immune system by histocompatibility antigen II (MCHII). Antigen fragments presented by normal cells, due to immune tolerance, do not elicit an immune response. Once normal cells become cancerous, the mutated gene expresses an aberrant protein, a fragment of which is presented on the cell surface, which causes a targeted immune response in the human immune system. Therefore, analysis of changes in BCR or TCR enables detection of tumor development and progression.
The invention has the beneficial effects that:
1. in the invention, 1300 samples of a control group of non-pancreatic cancer and TCR high-throughput sequencing data of 6 pancreatic cancer patients are firstly utilized to establish an artificial intelligence analysis model, and whether a patient with higher pancreatic cancer risk exists in a sample to be detected can be clearly judged by comparing with the pancreatic cancer specific TCR sequences.
2. The early pancreatic cancer can be found by analyzing TCR change through high-throughput sequencing, and the response of T cells in the human immune system to the pancreatic cancer is analyzed by utilizing the TCRCDR3 sequence specific to the pancreatic cancer, so that the method is a novel detection method.
3. By adopting a high-throughput sequencing technology, the invention can simultaneously compare a great number of specific TCR sequences, and has higher specificity and accuracy compared with the single detection of one or more markers.
4. The high-throughput sequencing instrument used in the invention has lower cost than large-scale imaging equipment, can be outsourced to a third party, and in addition, the labor cost for sampling and processing is lower than the labor cost for simultaneously detecting various markers and is also lower than the labor cost for a large number of cytological detections, so the detection cost is greatly reduced; in addition, the invention only needs to adopt a small amount of peripheral blood, and the sampling is simple, convenient and safe.
5. The TCR CDR3 sequence disclosed by the invention can be used for immunotherapy of pancreatic cancer.
Drawings
FIG. 1 shows the CDR3 sequence of the control group and the pancreatic cancer characteristic sequence of the present invention. The abscissa represents the sequence order in which the CDR3 sequence of a specific amino acid combination is added to the control sequence set or the pancreatic cancer characteristic sequence set, and the ordinate represents the number of times that the sequence repeats in a sample CXA logarithmic value of; the immune map of a pancreatic cancer patient has pancreatic cancer characteristic sequences of multiple types and high repetition times, few pancreatic cancer characteristic sequences of healthy people exist, and the pancreatic cancer characteristics of unknown subjects are obvious, which indicates that the risk of pancreatic cancer is high.
FIG. 2 is a graph showing that, in the present invention, pancreatic cancer characteristic indexes of healthy people, non-tumor patients, non-pancreatic cancer tumor patients and pancreatic cancer patients are calculated for a pancreatic cancer characteristic sequence set, and the pancreatic cancer characteristic indexes of healthy people, non-tumor patients and non-pancreatic cancer tumor patients are all significantly different from pancreatic cancer patients, which proves the specificity of the pancreatic cancer characteristic sequence set. Therefore, whether the unknown subject suffers from pancreatic cancer can be judged.
Detailed Description
The following description of the embodiments of the present invention is provided to facilitate the understanding of the present invention by those skilled in the art, but it should be understood that the present invention is not limited to the scope of the embodiments, and it will be apparent to those skilled in the art that various changes may be made without departing from the spirit and scope of the invention as defined and defined in the appended claims, and all matters produced by the invention using the inventive concept are protected.
Example 1 acquisition of the set of sequences of the TCR marker CDR3 of pancreatic cancer by Immunochromatographic analysis
1. Sampling and immune mapping (method reference patent ZL201910300069.9)
Collecting 1301 control groups (including healthy people and non-tumor disease patients, 1300 people for establishing a model, 1 healthy person for verifying), 7 pancreatic cancer patients (6 people for establishing a model, 1 person for verifying) and peripheral blood (10 mL per person) of 1 unknown health condition subject, obtaining the antigenic determinant 3(CDR3) amino acid sequences of the TCRs of the subject and the control groups by high-throughput sequencing, and ensuring that the total number of the CDR3 sequences of the functional TCRs of each sample is not less than 30000 comprehensively;
2. the CDR3 sequences of the TCR of each sample were randomly non-back sampled such that the sum of the number of CDR3 sequences of each sample was 30000. For any particular CDR3 sequence X, the number of repeats in a single sample sequencing result was counted as CX；
3. By analyzing the TCR CDR3 data, the pancreatic cancer TCR marker CDR3 sequence was determined:
a) summarizing and de-duplicating all CDR3 sequences of 1300 control group samples for establishing a model, and setting the CDR3 sequences as a control sequence set;
b) all CDR3 sequences of 6 pancreatic cancer samples used for modeling were summed up and de-duplicated, and then all sequences containing sequence repeats in the control sequence set were removed to prepare a pancreatic cancer characteristic sequence set. The plot is shown in FIG. 1A, in which the CDR3 sequence with the abscissa representing a specific amino acid combination is added to the sequence of the control sequence set or the pancreatic cancer characteristic sequence set, and the ordinate represents the number of times C that the sequence repeats in a sampleXThe logarithmic value of (c).
c) The immune profiles of 1 healthy human, 1 pancreatic cancer patient and 1 subject of unknown health status were mapped according to the same mapping method, referring to the control sequence set and the pancreatic cancer signature sequence set, see FIGS. 1B-D. As can be seen from the figure, the immune map of the pancreatic cancer patient contains more types of pancreatic cancer characteristic sequences with higher repeated occurrence frequency; in the immune map of a healthy person, only a few pancreatic cancer characteristic sequences exist; a subject with unknown health condition has a pancreatic cancer signature sequence higher than that of a healthy person, indicating that the person is at higher risk for pancreatic cancer.
d) Centralizing pancreatic cancer characteristic sequences, and repeating the times C of all CDR3 sequences in the single sample of the sequence participating in modeling pancreatic cancer samplesXThe total sum × of the sequences comprises the sequences, the number of samples participating in modeling pancreatic cancer is ranked from high to low, the top 100 are the pancreatic cancer TCR marker CDR3 sequences, and the specific sequences are shown in SEQ ID NO. 1-100.
Example 2 validation of the specificity of the pancreatic cancer TCR marker CDR3 sequence set
1. Sampling and immune mapping (method reference patent ZL201910300069.9)
Collecting peripheral blood (10 mL per person) of 349 tumor patients with non-pancreatic cancer and 2 subjects with unknown health conditions, and obtaining the amino acid sequences of antigenic determinant 3(CDR3) of TCR of the subjects and the control group by high-throughput sequencing to ensure that the total number of CDR3 sequences of functional TCR of each sample is not less than 30000 comprehensively; the CDR3 sequences of the TCR of each sample were randomly non-back sampled such that the sum of the number of CDR3 sequences of each sample was 30000.
2. 100 healthy and 45 patients with non-tumor diseases were randomly selected from the control group of example 1.
3. Pancreatic cancer characteristic indices were analyzed based on the immune profiles of 100 healthy persons, 45 non-neoplastic disease patients, 6 pancreatic cancer patients from example 1, and 349 newly acquired non-pancreatic cancer tumor patients, 2 unknown health condition subjects from example 2.
Wherein the pancreatic cancer characteristic index is defined as: in a sample, all CDR3 sequences belonging to the pancreatic cancer feature sequence set repeat within the sample by the number of times CXThe sum of (a) and (b). The results of the analysis are shown in Table 2 below and FIG. 2. The pancreatic cancer group has significant differences with healthy people (p ═ 1.9E-78), non-tumor diseases (p ═ 6.6E-43) and other tumors (p ═ 9.9E-162), which proves the specificity of the pancreatic cancer characteristic sequence set.
TABLE 2 pancreatic cancer characteristic index for different sample groups
4. Each group was analyzed for pancreatic cancer characteristic index (table 3), and among the subjects with unknown health conditions (test samples), 2 persons had pancreatic cancer characteristic index higher than the mean value of "other tumors" group +2 × SD (16.1+2 × 41.1 ═ 98.2), and these 2 persons had higher risk of pancreatic cancer. After comparison with the results of clinical examinations, this 2 persons were confirmed as pancreatic cancer patients. This example demonstrates the feasibility of using a pancreatic cancer signature set and a pancreatic cancer signature index to predict a subject's risk of developing pancreatic cancer.
TABLE 3 pancreatic cancer characteristic index analysis
| Healthy person | Non-neoplastic disease | Other tumors | Pancreatic cancer | Test sample | |
| Mean | 11.89 | 10.42 | 16.06 | 10978.67 | 2365.00 |
| SD | 11.90 | 11.97 | 41.08 | 2665.46 | 202.23 |
| mean+2SD | 35.69 | 34.36 | 98.23 |
In conclusion, the pancreatic cancer TCR marker CDR3 sequence of the present invention has significant pancreatic cancer specificity, and can be used not only for predicting pancreatic cancer risk of a subject, but also for biological immunotherapy of pancreatic cancer in the future.
Sequence listing
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<213> Artificial Sequence (Artificial Sequence)
<400>26
Ala Ser Ser Leu Val Pro Thr Ala Glu Gln Tyr
1 5 10
<210>27
<211>15
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>27
Ala Ser Ser Phe Ser Gly Gly Ser Gln Ser Thr Asp Thr Gln Tyr
1 5 10 15
<210>28
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>28
Ala Ser Lys Val Glu Gly Ile Thr Glu Ala Phe
15 10
<210>29
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>29
Gln Gln Phe Ile Gln Gly Cys Gly Asp Pro Val
1 5 10
<210>30
<211>7
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>30
Ser Thr Asp Glu Arg Gly Phe
1 5
<210>31
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>31
Ala Ser Ser Ala Pro Gly Thr Gly Trp His Glu Gln Phe
1 5 10
<210>32
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>32
Ala Ser Ser Leu Leu Gln Thr Gly Gly Leu Asp Glu Gln Tyr
1 5 10
<210>33
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>33
Ala Ser Ser Leu Val Gln Gly Thr Leu Ser Tyr Glu Gln Tyr
1 5 10
<210>34
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>34
Ala Ser Ser Pro Gln Pro Ala Val Lys Thr Gln Tyr
1 5 10
<210>35
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>35
Ala Ser Ser Pro Asn Asp Arg Val His Thr Ile Tyr
1 5 10
<210>36
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>36
Ala Ser Ser Leu Val Gly Leu Ala Gly Gly Ala Gln Tyr
1 5 10
<210>37
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>37
Ser Ala Arg Val Arg Glu Ala Asn Glu Gln Phe
1 5 10
<210>38
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>38
Ala Ser Ser Leu Leu Gly Phe Ser His Tyr Gly Tyr Thr
1 5 10
<210>39
<211>16
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>39
Ala Ser Ser Leu Leu Thr Gly Gly Ala Gly Ile Tyr Asn Glu Gln Phe
1 5 10 15
<210>40
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>40
Ala Ser Ser Ser Gln Ala Gly Val Asp Lys Thr Gln Tyr
1 5 10
<210>41
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>41
Ala Ser Thr Thr Asp Gly Arg Glu Thr Gln Tyr
1 5 10
<210>42
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>42
Ala Ser Lys Ser Trp Thr Ser Gln Glu Thr Gln Tyr
1 5 10
<210>43
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>43
Ala Ser Ser Leu Asn Ser Gly Gln Gly Ala Glu Thr Gln Tyr
1 5 10
<210>44
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>44
Ala Ser Ser His Trp Ala Gly Lys Ser Gly Asn Thr Ile Tyr
1 5 10
<210>45
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>45
Ala Ser Ser Leu Glu Gly Gln Arg Glu Pro Gln His
1 5 10
<210>46
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>46
Ala Ser Ser Pro Ser Thr Thr Gly Asp Thr Glu Ala Phe
1 5 10
<210>47
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>47
Gln Gln Phe Arg Pro Lys Thr Arg Leu Arg Ala Val
1 5 10
<210>48
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>48
Ala Ser Ser Leu Tyr Pro Gly Gln Gly Lys Ala Phe
1 5 10
<210>49
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>49
Ala Ser Ser Leu Val Gln Gly Ile Glu Glu Gln Tyr
1 5 10
<210>50
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>50
Ala Ser Ser Ser Gly Gln Asp Tyr Gly Gln Tyr
1 5 10
<210>51
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>51
Ala Ser Ser Glu Ser Arg Thr Gly Leu Asn Thr Ile Tyr
1 5 10
<210>52
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>52
Ala Ser Ser Leu Leu Gly Leu Glu Arg Asp Thr Gln Tyr
1 5 10
<210>53
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>53
Ala Ser Ser Leu Leu Arg Pro Gly Ser Ser Glu Gln Tyr
1 5 10
<210>54
<211>17
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>54
Ala Ser Ser Leu Ser Phe Gly Ala Thr Gly Asn Gly Gln Glu Thr Gln
1 5 10 15
Tyr
<210>55
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>55
Ala Ser Ser Pro Leu Leu Arg Asp Tyr Asn Glu Gln Phe
1 5 10
<210>56
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>56
Ala Gln Gly Gly Arg Gly Ser Tyr Gly Tyr Thr
1 5 10
<210>57
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>57
Ala Ser Thr Gln Asp Trp Gly Thr Gly Glu Leu Phe
1 5 10
<210>58
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>58
Ser Ala Leu Arg Arg Gly Gln Arg Asn Ser Pro Leu His
1 5 10
<210>59
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>59
Ala Ser Ser Leu Glu Ser Ala Thr Gln Ala Phe
1 5 10
<210>60
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>60
Ala Ser Ser Lys Leu Thr Pro Thr Glu Pro Gln His
1 5 10
<210>61
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>61
Ala Ser Ser Val Ser Pro Asp Pro Ser Gly Asn Thr Ile Tyr
1 5 10
<210>62
<211>10
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>62
Ala Ser Thr Phe Ser Gly Val Glu Gln Phe
1 5 10
<210>63
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>63
Ala Ser Ser Ser Gly Gln Asp Tyr Glu Arg Tyr
1 5 10
<210>64
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>64
Ala Ser Ser Asp Arg Ser Gly Tyr Pro Tyr Glu Gln Tyr
1 5 10
<210>65
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>65
Ala Ser Ser Asn Arg Asp Arg Asp Ser Ser Glu Gln Tyr
1 5 10
<210>66
<211>16
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>66
Ala Ser Ser Leu Glu Gly Thr Ser Gly Arg Asn Phe Asn Thr Gln Tyr
1 5 10 15
<210>67
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>67
Ala Ser Ser Leu Ala Leu Thr Gly Ser His Pro Glu Ser
1 5 10
<210>68
<211>15
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>68
Ala Ser Ser Tyr Ser Val Thr Gly Ile Leu Thr Gly Glu Leu Phe
1 5 10 15
<210>69
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>69
Ala Ser Ser Ser Gly Gln Asp Tyr Glu Gln Cys
1 5 10
<210>70
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>70
Ala Ser Ser Pro Ala Gly Ala Ser Gly Thr Glu Gln Phe
1 5 10
<210>71
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>71
Ala Ser Ser Pro Gly Pro Lys Ser Arg Tyr Glu Gln Tyr
1 5 10
<210>72
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>72
Ser Ala Arg Asp Gln Ala Thr Leu Val Asp Glu Gln Tyr
1 5 10
<210>73
<211>10
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>73
Ala Ser Arg Arg Asp Gly Glu Gly Tyr Thr
1 5 10
<210>74
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>74
Ala Ser Ser Leu Gly Thr Met Tyr Thr Glu Ala Phe
1 5 10
<210>75
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>75
Ala Ser Ile Arg Ser Gly Val Arg Tyr Asn Glu Gln Phe
1 5 10
<210>76
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>76
Ala Ser Ser Arg Pro Arg Ala Ser Gly Ser Tyr Glu Gln Tyr
1 5 10
<210>77
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>77
Ala Ser Ser Asp Ala Pro Gly Gly Ala Leu Trp Glu Gln Tyr
1 5 10
<210>78
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>78
Ala Ser Ser Ser Gln Arg Glu Lys Glu Thr Gln Tyr
1 5 10
<210>79
<211>16
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>79
Ala Ser Ser Leu Val Pro Arg Thr Gly Leu Ser Ser Tyr Glu Gln Tyr
1 5 10 15
<210>80
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>80
Ala Ser Ser Val Glu Ala Asp Ser Trp Asn Ser Pro Leu His
1 5 10
<210>81
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>81
Ala Ser Ser Thr Pro Thr Ser Asp Leu Pro Tyr Glu Gln Tyr
1 5 10
<210>82
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>82
Ala Ser Ser Glu Gln Gly Ala Ile Ser Tyr Gly Tyr Thr
15 10
<210>83
<211>16
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>83
Ala Ser Ser Leu Ala Gly Gln Gly Glu Ser Tyr Asn Tyr Gly Tyr Thr
1 5 10 15
<210>84
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>84
Ala Ser Ser Ser Lys Gly Pro Thr Gly Thr Asn Glu Gln Phe
1 5 10
<210>85
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>85
Ala Thr Gly Ala Gly Thr Gly Ala Ser Pro Leu His
1 5 10
<210>86
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>86
Ala Ser Gly Phe Gly Thr Glu Phe Tyr Glu Gln Tyr
1 5 10
<210>87
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>87
Ala Ser Ser Asn Asn Arg Gly Arg Asn Glu Lys Leu Phe
1 5 10
<210>88
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>88
Ala Ser Ser Tyr Asp Arg Ile Leu Thr Asp Thr Gln Tyr
1 5 10
<210>89
<211>13
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>89
Ala Ser Ser Leu Ala Ala Ala Ala Gly Tyr Gly Tyr Thr
1 5 10
<210>90
<211>11
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>90
Pro Ala Arg Gln Gly Glu Glu Arg Pro Ser Thr
1 5 10
<210>91
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>91
Ser Val Gly Ala Ala Gln Gly Asp Thr Glu Ala Phe
1 5 10
<210>92
<211>12
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>92
Ala Ser Arg Gln Gly Gln Gly Thr Ser Pro Leu His
1 5 10
<210>93
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>93
Ala Ser Ser Leu Asn Pro Gly Arg Pro Thr Tyr Glu Gln Tyr
1 5 10
<210>94
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>94
Ala Ser Ser Arg Arg Gln Ala Leu Ser Thr Asp Thr Gln Tyr
1 5 10
<210>95
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>95
Ala Ser Ser Leu Asp Arg Gly Gln Ile Gly Thr Glu Ala Phe
1 5 10
<210>96
<211>10
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>96
Ala Ser Ser Thr His Pro Arg Ser Gln Tyr
1 5 10
<210>97
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>97
Ala Ser Ser Tyr Gly Gln Gly Lys Met Asn Thr Glu Ala Phe
1 5 10
<210>98
<211>15
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>98
Ala Ser Ser Leu Asp Leu Gly Leu Val Asn Thr Gly Glu Leu Phe
1 5 10 15
<210>99
<211>14
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>99
Ala Ser Ser Phe Gly Pro Val Gln Ala Asn Tyr Gly Tyr Thr
1 5 10
<210>100
<211>15
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>100
Ala Ser Ser Ser Leu Phe Pro Gly Gln Gly Arg Gly Glu Gln Phe
1 5 10 15
Claims (7)
1. A peripheral blood TCR marker of pancreatic cancer, which is characterized by comprising at least one protein shown as SEQ ID NO. 1-100.
2. The peripheral blood TCR marker for pancreatic cancer according to claim 1, wherein the protein sequence of the marker is a protein which can express the same function after one or more bases are substituted, deleted and/or substituted by the sequence shown in SEQ ID No. 1-100.
3. Use of a marker according to claim 1 for the preparation of a formulation for the treatment of pancreatic cancer.
4. The use of claim 3, wherein the preparation comprises a T cell receptor comprising the marker, or a plasmid, viral vector or nucleic acid fragment capable of expressing the T cell receptor producing the marker.
5. A kit for detecting pancreatic cancer, comprising an antibody that specifically binds to the marker of claim 1.
6. An agent comprising an antibody that specifically binds to the marker of claim 1.
7. A protein chip for detecting pancreatic cancer, comprising a substrate and a specific antibody spotted on the substrate, wherein the specific antibody is an antibody capable of specifically binding to the marker of claim 1.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010665297.9A CN111624340A (en) | 2020-07-11 | 2020-07-11 | Peripheral blood TCR marker of pancreatic cancer and detection kit and application thereof |
| PCT/CN2021/101981 WO2022012292A1 (en) | 2020-07-11 | 2021-06-24 | Peripheral blood tcr marker for pancreatic cancer, and detection kit and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010665297.9A CN111624340A (en) | 2020-07-11 | 2020-07-11 | Peripheral blood TCR marker of pancreatic cancer and detection kit and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111624340A true CN111624340A (en) | 2020-09-04 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010665297.9A Pending CN111624340A (en) | 2020-07-11 | 2020-07-11 | Peripheral blood TCR marker of pancreatic cancer and detection kit and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN111624340A (en) |
| WO (1) | WO2022012292A1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113109564A (en) * | 2021-03-15 | 2021-07-13 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof |
| CN113567682A (en) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of Alzheimer disease and detection kit and application thereof |
| WO2022012292A1 (en) * | 2020-07-11 | 2022-01-20 | 成都益安博生物技术有限公司 | Peripheral blood tcr marker for pancreatic cancer, and detection kit and use thereof |
| WO2022194039A1 (en) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | Peripheral blood tcr marker for acute b lymphocytic leukemia, and detection kit thereof and use thereof |
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| CN107299110B (en) * | 2017-05-27 | 2019-11-22 | 上海优卡迪生物医药科技有限公司 | A CAR-T therapeutic vector for pancreatic cancer and malignant mesothelioma based on OCTS technology and its construction method and application |
| WO2020028671A1 (en) * | 2018-08-03 | 2020-02-06 | Scripps Health | Detection and isolation of myeloid-derived suppressor cell subpopulations |
| CN111402957B (en) * | 2020-03-10 | 2021-05-18 | 成都益安博生物技术有限公司 | Immune characteristic recognition method based on neural network |
| CN111624340A (en) * | 2020-07-11 | 2020-09-04 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of pancreatic cancer and detection kit and application thereof |
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2020
- 2020-07-11 CN CN202010665297.9A patent/CN111624340A/en active Pending
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2021
- 2021-06-24 WO PCT/CN2021/101981 patent/WO2022012292A1/en active Application Filing
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022012292A1 (en) * | 2020-07-11 | 2022-01-20 | 成都益安博生物技术有限公司 | Peripheral blood tcr marker for pancreatic cancer, and detection kit and use thereof |
| CN113109564A (en) * | 2021-03-15 | 2021-07-13 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof |
| WO2022194039A1 (en) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | Peripheral blood tcr marker for acute b lymphocytic leukemia, and detection kit thereof and use thereof |
| CN113567682A (en) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of Alzheimer disease and detection kit and application thereof |
| WO2023000688A1 (en) * | 2021-07-23 | 2023-01-26 | 成都益安博生物技术有限公司 | Peripheral blood tcr marker of alzheimer's disease, and detection kit and application thereof |
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| Publication number | Publication date |
|---|---|
| WO2022012292A1 (en) | 2022-01-20 |
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