CN113109564A - Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof - Google Patents
Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof Download PDFInfo
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- CN113109564A CN113109564A CN202110273700.8A CN202110273700A CN113109564A CN 113109564 A CN113109564 A CN 113109564A CN 202110273700 A CN202110273700 A CN 202110273700A CN 113109564 A CN113109564 A CN 113109564A
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57426—Specifically defined cancers leukemia
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57488—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds identifable in body fluids
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- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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Abstract
The invention discloses a TCR marker of peripheral blood of acute myelocytic leukemia and 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 through high-throughput sequencing and TCR data analysis, a characteristic TCR sequence in the peripheral blood of the acute myelocytic leukemia is determined firstly, and then a test result of a sample to be tested is compared with the characteristic TCR sequence, so that whether the acute myelocytic leukemia is suffered or not is determined. The invention can simultaneously compare a great number of acute myelocytic leukemia 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 acute myelocytic leukemia, a detection kit and application thereof.
Background
Acute Myelogenous Leukemia (AML) is a Myeloid hematopoietic stem/progenitor malignancy. The abnormal hyperplasia of primary and juvenile medullary cells in bone marrow and peripheral blood is taken as a main characteristic, the clinical manifestations are anemia, bleeding (thrombocytopenia), infection and fever, organ infiltration, skin infiltration, abnormal metabolism, gingival hyperplasia and the like, most cases are urgent and serious, the disease condition progresses rapidly, the natural course of disease is only from several weeks to several months, the prognosis is extremely dangerous, and the life can be threatened if the treatment is not performed in time. The disease accounts for 30% of the leukemia of children. Childhood acute myelogenous leukemia is similar to adults (<50 years) in molecular biological changes and chemotherapy response. Acute myeloid leukemia in infants is more prone to extramedullary leukemia than adults. Acute leukemia is more common than chronic leukemia in China, and the most acute myelocytic leukemia is (1.62/10 ten thousand).
At present, the diagnosis means aiming at the acute myelocytic leukemia mainly comprises blood routine detection, cytochemical examination, immunophenotyping, chromosome detection and bone marrow image detection.
Anemia and thrombocytopenia are very common (accounting for 75-95%) in routine blood tests, and the white blood cell count of half acute myelocytic leukemia patients is increased by 10 multiplied by 109/L～100×109between/L, some cases even>100×109And L. The white blood cell count of some patients is normal, and the white blood cell count of a few patients (usually M3 type or old age cases)<4.0×109And L. Some patients have lower than normal hemoglobin, even have severe anemia, and have a decrease in reticulocytes. The platelet count is reduced in most patients and increased in a few patients, normally or mildly. Such tests may be high or low depending on the patient and may not be used to characterize acute myelogenous leukemia.
The cytochemical staining characteristics of different subtypes of acute myelocytic leukemia are different, so that the cytochemical staining of the acute myelocytic leukemia is very important for diagnosing the disease. Common methods of cytochemical staining include myeloperoxidase staining (MPO), Sudan black B Staining (SBB), Naphthol acetate AS-D esterase staining (NAS-DCE), Naphthol butyrate staining (alpha-NBE), Naphthol acetate esterase staining (alpha-NAE), periodate-Schiff staining (glycogen staining, PAS), acid phosphatase staining (ACP), alkaline phosphatase staining (NAP), lysozyme and the like. If necessary, esterase double staining and Phl (phi) corpuscle can be carried out. This test cannot be used for the early diagnosis of acute myeloid leukemia.
The immune classification is to determine the series sources according to the related antigens expressed by the acute myelocytic leukemia cells, and to further classify the acute myelocytic leukemia into different subtypes by analyzing with a multi-parameter flow cytometer. The sensitivity of the detection is not high due to the influence of the amount of the relevant antigen.
80-90% of childhood acute myeloid leukemia is associated with chromosomal abnormalities. Of these, about half of cases of acute myelogenous leukemia appear as a single karyotype abnormality, with the remainder being accompanied by additional abnormalities. By adopting a high-resolution technology, the discovery rate of the karyotype abnormality is up to more than 90%. Chromosomal abnormalities in acute myeloid leukemia are dominated by structural abnormalities, as many as 39, and certain specific structural abnormalities, such as t (8; 21) (q 22; q22), t (15; 17) (q 22; q21) and inv (16) (p 13; q22) or t (16; 16) (p 13; q11), are associated with a good prognosis. Such detection is not unique enough to be used for early diagnosis.
Myelograms detect high proliferation in most patients, and normal hematopoietic cells are replaced by leukemia cells; a few patients have low myeloproliferation, but the number of primitive cells is still above 30%. The discovery of Auer bodies in the cytoplasm is more helpful to eliminate Acute Lymphocytic Leukemia (ALL) and confirm the diagnosis of acute myelogenous leukemia this test can be high or low depending on the patient's condition and cannot be used as a qualitative test for acute myelogenous leukemia.
In addition to the above tests, there are very few markers currently available for diagnosing acute myeloid leukemia.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides the TCR marker of the peripheral blood of the acute myelocytic leukemia and the detection kit and the application thereof, which can accurately and quickly judge whether a sample to be detected has a patient with higher acute myelocytic leukemia risk.
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 acute myelocytic leukemia comprises at least one of proteins with sequences shown in SEQ ID NO. 1-100, and the specific sequences are 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 acute myelocytic leukemia.
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 detecting acute myelogenous leukemia, comprising an antibody capable of specifically binding to the marker.
An agent comprising an antibody that specifically binds to the marker; the preparation can be used for diagnosing, predicting, detecting or screening acute myelogenous leukemia.
A protein chip for detecting acute myelogenous leukemia comprises a substrate and specific antibodies which are spotted on the substrate, wherein the specific antibodies are antibodies which can be specifically combined with the markers.
According to the inventionThe principle is as follows: b-lymphocytes and T-lymphocytes in the human body are two important types of cells in the adaptive immune system. B cells recognize antigens through a B Cell Receptor (BCR) on the cell surface, and later, BCR expresses antibodies and is secreted extracellularly when B cells differentiate 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-acute myelocytic leukemia and TCR high-throughput sequencing data of 45 patients with acute myelocytic leukemia are used for establishing an artificial intelligent analysis model, and whether a sample to be detected has a higher risk of acute myelocytic leukemia can be clearly judged by comparing with the specific TCR sequences of the acute myelocytic leukemia;
2. the early acute myelocytic leukemia can be found by analyzing the TCR change through high-throughput sequencing, and the T cell response to the acute myelocytic leukemia in the human immune system is analyzed by utilizing the specific TCR CDR3 sequence of the acute myelocytic leukemia, so that the method is a novel detection method;
3. the invention adopts high-throughput sequencing technology to simultaneously compare a great number of specific TCR sequences, and has higher specificity and accuracy compared with the method of singly detecting 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;
5. the invention only needs to adopt a small amount of peripheral blood, and the sampling is simple, convenient and safe;
6. the TCR CDR3 sequence disclosed by the invention can be used for immunotherapy of acute myelocytic leukemia.
Drawings
FIG. 1 shows the discovery of TCR sequences characteristic of acute myelogenous leukemia using an immune-based big data analysis system in accordance with the present invention. The abscissa represents the sequence order of the CDR3 sequence of a specific amino acid combination added to the control sequence set or the acute myelocytic leukemia characteristic sequence set, and the ordinate represents the repeated occurrence number C of the sequence in a sampleXA logarithmic value of; the immune map of the acute myelocytic leukemia patient has a plurality of types of acute myelocytic leukemia characteristic sequences with high repetition times, few acute myelocytic leukemia characteristic sequences of healthy people are available, the acute myelocytic leukemia characteristics of unknown subjects are obvious, and the risk of suffering from acute myelocytic leukemia is high.
FIG. 2 is a graph showing the comparative analysis of acute myelogenous leukemia and other diseases by using the characteristic index of acute myelogenous leukemia in the present invention. The acute myelocytic leukemia characteristic indexes of healthy people, non-tumor patients and non-acute myelocytic leukemia tumor patients are obviously different from those of acute myelocytic leukemia patients, and the specificity of the acute myelocytic leukemia characteristic sequence set is proved. Therefore, whether the unknown subject suffers from acute myelogenous leukemia 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 acute myelogenous leukemia TCR marker CDR3 sequence set obtained by immune map analysis
1. Sampling and immune profiling
Collecting 1301 control groups (including healthy people and non-tumor disease patients, 1300 people are used for establishing a model, 1 healthy person is used for verifying), 41 acute myelocytic leukemia patients (40 people are used for establishing a model, 1 person is used for verifying) and peripheral blood (10 mL per person) of 1 unknown health condition test subject, obtaining the amino acid sequences of the antigenic determinant 3(CDR3) of the TCR of the test subject and the control group by high-throughput sequencing, and ensuring that the total number of the CDR3 sequences of the functional TCR of each sample is not less than 25000 comprehensively;
2. the total number of CDR3 sequences for each functional TCR summed over the sequence of the sample 30000 was randomly sampled without back sampling to give a total of 30000 CDR3 sequences for that sample. All samples thus far contained a total number of CDR3 sequences for functional TCRs of 25000-30000. 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 acute myelocytic leukemia 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 40 acute myelocytic leukemia samples used for establishing the model are summed up and de-duplicated, and then all sequences containing sequence repeats in the comparison sequence set are removed to set the characteristic sequence set of the acute myelocytic leukemia. The graph is shown in FIG. 1A, wherein the abscissa represents the sequence of CDR3 with a specific amino acid combination added to the sequence set of control sequences or the sequence set of acute myelogenous leukemia characteristic sequences, and the ordinate represents the number of times C that the sequence repeats in a sampleXThe logarithmic value of (c).
c) According to the same mapping method, the immune maps of 1 healthy person, 1 acute myelogenous leukemia patient and 1 subject with unknown health condition are mapped with reference to the reference sequence set and the acute myelogenous leukemia characteristic sequence set, as shown in FIGS. 1B-D. As can be seen, the immune map of the patient with acute myelogenous leukemia contains more types of acute myelogenous leukemia characteristic sequences with higher repeated occurrence number (FIG. 1B); in the immune map of healthy people, there are only a few acute myelocytic leukemia characteristic sequences (fig. 1C); the subjects with unknown health had a higher signature sequence for acute myelogenous leukemia than healthy persons, indicating that the persons were at higher risk for acute myelogenous leukemia (FIG. 1D).
d) Centralizing the characteristic sequences of the acute myelocytic leukemia, and repeating the times of repeated appearance in a single sample of all the CDR3 sequences in two or more samples participating in modeling the acute myelocytic leukemiaXThe total number of the sequences is multiplied by the number of samples which contain the sequences and participate in modeling acute myelocytic leukemia, the samples are ranked from high to low, the first 100 are the TCR marker CDR3 sequences of the acute myelocytic leukemia, and the specific sequences are shown in SEQ ID NO. 1-100.
Example 2 validation of the specificity of the CDR3 sequence set of the TCR marker of acute myelogenous leukemia
1. Sampling and immune profiling
Collecting 435 tumor patients with non-acute myelogenous leukemia and peripheral blood (10 mL per person) of 2 subjects with unknown health conditions, and obtaining the antigenic determinant 3(CDR3) amino acid sequences of TCRs of the subjects and the control group by high-throughput sequencing to ensure that the total number of CDR3 sequences of the functional TCRs of each sample is not less than 25000 comprehensively; the total number of CDR3 sequences for each functional TCR summed over the sequence of the sample 30000 was randomly sampled without back sampling to give a total of 30000 CDR3 sequences for that sample. All samples thus far contained a total number of CDR3 sequences for functional TCRs of 25000-30000.
2. 100 healthy and 43 patients with non-tumor diseases were randomly selected from the control group of example 1.
3. Acute myelogenous leukemia characteristic indices were analyzed based on the immune profiles of 100 healthy patients, 43 non-neoplastic disease patients, 40 acute myelogenous leukemia patients from example 1, and 435 non-acute myelogenous leukemia tumor patients newly obtained in example 2, 2 subjects of unknown health status.
Wherein the acute myeloid leukemia characteristic index is defined as: in a sample, all CDR3 sequences belonging to the characteristic sequence set of acute myelogenous leukemia repeat within the sample by number CXThe sum of (a) and (b). The results of the analysis are shown in Table 2 below and FIG. 2. The acute myelocytic leukemia group has obvious difference with healthy people (p ═ 1.3E-75), non-tumor diseases (p ═ 5.4E-40) and other tumors (p ═ 4.8E-271), which proves the specificity of the characteristic sequence set of the acute myelocytic leukemia.
TABLE 2 characteristic indices of acute myelogenous leukemia for different sample groups
4. The acute myelogenous leukemia characteristic index of each group was analyzed (table 3), and the acute myelogenous leukemia characteristic index of 2 subjects with unknown health conditions (test samples) was significantly higher than the mean value of the "other tumors" group +2 × SD (67.8+2 × 258.3 ═ 584.4), and this 2 persons had a higher risk of suffering from acute myelogenous leukemia. After comparison with the clinical examination results, these 2 patients were identified as early-stage acute myelocytic leukemia patients. This example demonstrates the feasibility of predicting a subject's risk of developing acute myeloid leukemia using the acute myeloid leukemia signature set and the acute myeloid leukemia signature index.
TABLE 3 index analysis of acute myelogenous leukemia characteristics
In conclusion, the TCR marker CDR3 sequence of acute myelocytic leukemia of the present invention has significant specificity to acute myelocytic leukemia, and can be used not only for predicting the risk of acute myelocytic leukemia of the subject, but also for biological immunotherapy of acute myelocytic leukemia in the future.
Sequence listing
<110> Chengdu Yianbo Biotech Ltd
<120> acute myelocytic leukemia peripheral blood TCR marker, detection kit and application thereof
<160> 100
<170> SIPOSequenceListing 1.0
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<212> PRT
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Ala Thr Ser Arg Asp Leu Ala Gly Gly Val Gly Gly Tyr Thr
1 5 10
<210> 2
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Ser Val Val Thr Gly Gly Glu Val Tyr Glu Gln Tyr
1 5 10
<210> 3
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 3
Ala Ser Ser Pro Val Val Gly Asn Leu Asp
1 5 10
<210> 4
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ser Leu Arg Thr Asp Ser Ser Asn Glu Gln Phe
1 5 10
<210> 5
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 5
Ser Val Gly Thr Phe Trp Gly Gly Asp Glu Gln Phe
1 5 10
<210> 6
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 6
Ala Ser Ser Asp Glu Gly Gly Gly Pro Asp Thr Gln Tyr
1 5 10
<210> 7
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 7
Ala Thr Ser Glu Trp Asp Arg Ala His Glu Gln Phe
1 5 10
<210> 8
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ala Ile Ser Glu Ser Ile Thr Gly Leu Ala Tyr Glu Gln Tyr
1 5 10
<210> 9
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 9
Ser Val Ala Met Arg Gly Leu Asp Thr Asp Thr Gln Tyr
1 5 10
<210> 10
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 10
Ala Ser Ser Pro Lys Arg Asp Ser Glu Arg Ile Gln His
1 5 10
<210> 11
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 11
Ala Ser Ser His Pro Asp Arg Gly His Ser Tyr Glu Gln Tyr
1 5 10
<210> 12
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 12
Ala Ser Ser Pro Pro Val Val Asp Thr Ile Tyr
1 5 10
<210> 13
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 13
Ala Ser Ser Glu Asp Leu Lys Glu Pro Ala Val
1 5 10
<210> 14
<211> 13
<212> PRT
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<400> 14
Ala Ser Ser Leu Leu Arg Ser Thr Gly Val Glu Gln Phe
1 5 10
<210> 15
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 15
Ala Trp Ser Thr Gly Asp Phe Thr Gly Glu Leu Phe
1 5 10
<210> 16
<211> 13
<212> PRT
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<400> 16
Ala Ser Ser Ser Tyr Arg Arg Ala Gly Val Glu Gln Tyr
1 5 10
<210> 17
<211> 12
<212> PRT
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<400> 17
Ala Thr Ser Ile Gly Gly Pro Thr Asp Thr Gln Tyr
1 5 10
<210> 18
<211> 13
<212> PRT
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<400> 18
Ser Ala Arg Gln Ser Gly Arg Pro Tyr Asn Glu Gln Phe
1 5 10
<210> 19
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 19
Ser Ala Ser Ile Leu Gly Ser Lys Asn Gly Tyr Thr
1 5 10
<210> 20
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 20
Ala Ser Ser Trp Thr Gly Leu Val Gly Asp Gly Tyr Thr
1 5 10
<210> 21
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 21
Gln Gln Leu Leu Ser Thr Cys Arg Cys Arg Ala Val
1 5 10
<210> 22
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 22
Gln Gln Pro Val Val Arg Pro Thr Gly Leu Leu Arg Ala Val
1 5 10
<210> 23
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 23
Ala Ser Ile Leu Leu Leu Arg Gly Tyr Thr
1 5 10
<210> 24
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 24
Ala Ser Ser Leu Arg Glu Ser Ala Gly Lys Glu Thr Gln Tyr
1 5 10
<210> 25
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 25
Ala Thr Ser Ser Gln Glu Trp Ala Gln Phe
1 5 10
<210> 26
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 26
Ala Ser Tyr Asp Gly Ala Thr Asn Glu Lys Leu Phe
1 5 10
<210> 27
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 27
Ala Ser Ser Glu Asn Pro Asp Val Gly Tyr Gly Tyr Thr
1 5 10
<210> 28
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 28
Ala Ser Ser Pro Gly Leu Gly Gln Gly Asp Glu Gln Tyr
1 5 10
<210> 29
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 29
Ser Ala Pro Gly Leu Val Leu Gly Glu Gln Tyr
1 5 10
<210> 30
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 30
Ala Ser Ser Pro Ser Leu Ser Glu Ser Pro Asp Thr Gln Tyr
1 5 10
<210> 31
<211> 16
<212> PRT
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<400> 31
Ala Ser Ser Glu Ala Gln Val Ile Gly Gly Asn Thr Gly Glu Leu Phe
1 5 10 15
<210> 32
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 32
Ala Ile Ser Glu Ser Pro Arg Lys Ser Tyr Glu Gln Tyr
1 5 10
<210> 33
<211> 13
<212> PRT
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<400> 33
Ala Ser Ser Pro Pro Gly Gly Glu Tyr Tyr Glu Gln Tyr
1 5 10
<210> 34
<211> 13
<212> PRT
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<400> 34
Ala Trp Ser Asp Arg Gly Ala Phe Ser Asp Thr Gln Tyr
1 5 10
<210> 35
<211> 16
<212> PRT
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<400> 35
Ala Ser Ser Leu Asp Ala Gly Leu Trp Thr Gly Ala Asn Val Leu Thr
1 5 10 15
<210> 36
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 36
Ala Ser Ser Leu Leu Ser Leu Arg Glu Asn Thr Gly Glu Leu Phe
1 5 10 15
<210> 37
<211> 12
<212> PRT
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<400> 37
Ala Ser Thr Leu Gly Thr Ser Gly Arg Glu Gln Phe
1 5 10
<210> 38
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 38
Ser Ala Arg Asp Phe Arg Phe Gln Asp Gln Pro Gln His
1 5 10
<210> 39
<211> 11
<212> PRT
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<400> 39
Ala Ser Tyr Pro Pro Ala Ser Pro Arg Ala Phe
1 5 10
<210> 40
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 40
Ala Ser Ala Gly Pro Gly Gln Val Glu Ala Asp Thr Gln Tyr
1 5 10
<210> 41
<211> 12
<212> PRT
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<400> 41
Ser Ala Asp Gly Thr Ala Lys Gly Ser Pro Leu His
1 5 10
<210> 42
<211> 13
<212> PRT
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<400> 42
Ala Ser Ser Leu Asp Tyr Leu Val Asn Thr Glu Ala Phe
1 5 10
<210> 43
<211> 12
<212> PRT
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<400> 43
Ala Ser Ser Arg Glu Leu Met Ser Asn Glu Gln Phe
1 5 10
<210> 44
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 44
Ser Ala Gln Ala Gly Gly Leu Thr Tyr Glu Gln Tyr
1 5 10
<210> 45
<211> 13
<212> PRT
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<400> 45
Arg Glu Asn Gly Arg Pro Gly His Gln Glu Thr Gln Tyr
1 5 10
<210> 46
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 46
Ala Trp Arg Gly Asn Ser Tyr Gly Tyr Thr
1 5 10
<210> 47
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 47
Ser Ala Met Gly Thr Ser Gly Ala Thr Tyr Glu Gln Tyr
1 5 10
<210> 48
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 48
Ala Ser Ser Arg Thr Gly Gly Tyr Pro Tyr Glu Gln Tyr
1 5 10
<210> 49
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 49
Ala Thr Glu Lys Gly Thr Gly Thr Ala Ser Asn Gln Pro Gln His
1 5 10 15
<210> 50
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 50
Ala Ser Ser Ala Gly Gln Glu Leu Thr Glu Ala Phe
1 5 10
<210> 51
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 51
Ala Ser Leu Gly Arg Tyr Arg Pro Lys Met Asn Thr Glu Ala Phe
1 5 10 15
<210> 52
<211> 11
<212> PRT
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<400> 52
Ala Ser Ser Ser Val Gly Tyr Asn Thr Gln Tyr
1 5 10
<210> 53
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 53
Ala Ser Ser Ala Thr Gly Trp Glu Thr Gln Tyr
1 5 10
<210> 54
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 54
Ala Thr Ser Pro Gly Phe Gly Glu Gly Tyr Asn Glu Gln Phe
1 5 10
<210> 55
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 55
Ala Ile Ser Asp Ser Gly Thr Val Asn Thr Glu Ala Phe
1 5 10
<210> 56
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 56
Ala Ser Thr Thr Pro Gly Gly Ser Ser Tyr Asn Glu Gln Phe
1 5 10
<210> 57
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 57
Ala Ser Ser Gln Glu Trp Asp Arg Tyr Tyr Thr
1 5 10
<210> 58
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 58
Ala Ser Ser Pro Gln Gly Trp Val Asp Tyr Gly Tyr Thr
1 5 10
<210> 59
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 59
Ala Ser Ser Leu Glu Gly Gly Pro Ser Met Gln Glu Thr Gln Tyr
1 5 10 15
<210> 60
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 60
Ala Ser Ser Phe Lys Glu Leu Asn Thr Gly Glu Leu Phe
1 5 10
<210> 61
<211> 16
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 61
Ala Ser Ser Ser Pro Gly Gly Val Pro Ser Gly Ala Asn Val Leu Thr
1 5 10 15
<210> 62
<211> 12
<212> PRT
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<400> 62
Ala Thr Ser Asp Pro Gly Leu Pro Tyr Gly Tyr Thr
1 5 10
<210> 63
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 63
Ala Thr Ser Gly Ala Gly Gly Gln Gly Gly Glu Gln Tyr
1 5 10
<210> 64
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 64
Ser Ala Arg Gly Arg Leu Ala Thr Leu Arg Asp Thr Gln Tyr
1 5 10
<210> 65
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 65
Ala Thr Arg Val Arg Thr Gly His Asn Gln Pro Gln His
1 5 10
<210> 66
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 66
Ser Ala Lys Val Asn Ser Tyr Glu Gln Tyr
1 5 10
<210> 67
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 67
Ala Ile Ser Pro Val Val Gly Gly Ser Tyr Glu Gln Tyr
1 5 10
<210> 68
<211> 13
<212> PRT
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<400> 68
Ala Ser Ser Pro Ser Lys Pro Thr Pro Glu Thr Gln Tyr
1 5 10
<210> 69
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 69
Ala Ser Ser Pro Tyr Gly Gly Gly Ala Ser Gly Tyr Thr
1 5 10
<210> 70
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 70
Gln Gln Leu Val Ser Gly Arg Ala Arg Asp Pro Val
1 5 10
<210> 71
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 71
Ala Ser Ser Asp Ser Phe Gly Ala Ala Asp Thr Gln Tyr
1 5 10
<210> 72
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 72
Ala Ile Ser Thr Ser Ala Ser Asn Gln Pro Gln His
1 5 10
<210> 73
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 73
Ala Ile Ser Gly Thr Gly Ile Thr Ile Tyr
1 5 10
<210> 74
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 74
Ala Ile Ser Glu Ser Asn Ser Arg Asp Thr Gln Tyr
1 5 10
<210> 75
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 75
Ala Ser Ser His Met Thr Gly Asp Tyr Thr Tyr Asn Glu Gln Phe
1 5 10 15
<210> 76
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 76
Ser Ala Ile Gln Gly Val Ala Gly Ser Pro Leu His
1 5 10
<210> 77
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 77
Ala Ser Ser Asp Arg Phe Gly Lys Ile Gly Glu Gln Phe
1 5 10
<210> 78
<211> 16
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 78
Ser Ala Arg Asp Arg Trp Ala Ser Gly Phe Ser Thr Asp Thr Gln Tyr
1 5 10 15
<210> 79
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 79
Ser Val Pro Gly Pro Phe Asn Gln Pro Gln His
1 5 10
<210> 80
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 80
Ser Thr Arg Thr Gly Pro Leu Asn Glu Gln Phe
1 5 10
<210> 81
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 81
Ala Ser Ser Pro Glu Leu Gly Gly Asn Glu Lys Leu Phe
1 5 10
<210> 82
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 82
Ser Ala Pro Ala Arg Ala Asp Thr Glu Ala Phe
1 5 10
<210> 83
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 83
Ser Ala Ala Gly Leu Glu Gly Glu Thr Gln Tyr
1 5 10
<210> 84
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 84
Ser Ala Gly Gln Val Ala Gly Val Gly Glu Gln Tyr
1 5 10
<210> 85
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 85
Ala Ser Ser Pro Pro Ile Gly Trp Arg Glu Tyr Asn Glu Gln Phe
1 5 10 15
<210> 86
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 86
Ala Ser Ser Tyr Trp Gly Gly Glu Asn Asn Glu Gln Phe
1 5 10
<210> 87
<211> 8
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 87
Ser Val Ser Asp Val Thr Gln Tyr
1 5
<210> 88
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 88
Ala Trp Ser Val Gln Thr Thr Lys Ala Phe
1 5 10
<210> 89
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 89
Ala Ser Ser Pro Ser Pro Thr Gly Ala Thr Asp Thr Gln Tyr
1 5 10
<210> 90
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 90
Ala Ser Asn Glu Gly Gln Gly Ala Gly Glu Gln Tyr
1 5 10
<210> 91
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 91
Ser Ala Arg Ala Val Thr Gly Ala Asp Gln Pro Gln His
1 5 10
<210> 92
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 92
Ala Ser Ser Leu Leu Leu Ser Gly Arg Lys Asn Glu Gln Phe
1 5 10
<210> 93
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 93
Ala Ser Arg Lys Ser Gly Asp Arg Thr Asp Thr Gln Tyr
1 5 10
<210> 94
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 94
Ala Ser Thr Asn Arg Gly Ser Asn Thr Glu Ala Phe
1 5 10
<210> 95
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 95
Ala Ser Ser Pro Pro Gly Gly Val Gly Ser Asn Gln Pro Gln His
1 5 10 15
<210> 96
<211> 16
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 96
Ala Ser Ser Glu Gly Leu Phe Phe Gln Ser Ser Tyr Asn Glu Gln Phe
1 5 10 15
<210> 97
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 97
Ala Ser Ser Ala Gly Leu Gly Val Gln Tyr
1 5 10
<210> 98
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 98
Ser Ala Glu Glu Gly Gly Ile Ser Ile Tyr Asn Ser Pro Leu His
1 5 10 15
<210> 99
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 99
Ser Ala Arg Ser Gly Ala Ser Gly Arg Phe Ser Tyr Glu Gln Tyr
1 5 10 15
<210> 100
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 100
Ala Trp Ser Lys Ser Gly Ala Asn Glu Gln Phe
1 5 10
Claims (7)
1. A peripheral blood TCR marker of acute myelocytic leukemia, which is characterized by comprising at least one protein shown as SEQ ID NO. 1-100.
2. The TCR marker for peripheral blood of acute myelocytic leukemia 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 replaced 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 acute myeloid leukemia.
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 the detection of acute myeloid leukemia 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; the preparation can be used for diagnosing, predicting, detecting or screening acute myelogenous leukemia.
7. A protein chip for detecting acute myelogenous leukemia, which comprises a substrate and specific antibodies spotted on the substrate, wherein the specific antibodies are antibodies capable of specifically binding to the marker of claim 1.
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| CN202110273700.8A CN113109564A (en) | 2021-03-15 | 2021-03-15 | Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof |
| PCT/CN2022/080335 WO2022194042A1 (en) | 2021-03-15 | 2022-03-11 | Peripheral blood tcr marker for acute myeloid leukaemia, and test kit and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110273700.8A CN113109564A (en) | 2021-03-15 | 2021-03-15 | Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof |
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| CN113109564A true CN113109564A (en) | 2021-07-13 |
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| CN202110273700.8A Pending CN113109564A (en) | 2021-03-15 | 2021-03-15 | Peripheral blood TCR marker of acute myelocytic leukemia and detection kit and application thereof |
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| CN (1) | CN113109564A (en) |
| WO (1) | WO2022194042A1 (en) |
Cited By (3)
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| 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 |
| CN113563454A (en) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | IgA nephropathy peripheral blood TCR marker and detection kit and application thereof |
| WO2022194042A1 (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 myeloid leukaemia, and test kit and application thereof |
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| 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 |
| CN113563454A (en) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | IgA nephropathy peripheral blood TCR marker 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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| WO2022194042A1 (en) | 2022-09-22 |
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