CN116554331B - Anti-human CD15 engineering antibody and application thereof - Google Patents
Anti-human CD15 engineering antibody and application thereof Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/567—Framework region [FR]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides an anti-human CD15 engineering antibody and application thereof, wherein the engineering antibody comprises a heavy chain variable region and a light chain variable region; the complementarity determining regions of the heavy chain variable region include CDR1, CDR2 and CDR3; the amino acid sequence of CDR1 of the heavy chain variable region is shown as SEQ ID NO. 1; the amino acid sequence of CDR2 of the heavy chain variable region is shown as SEQ ID NO. 2; the amino acid sequence of CDR3 of the heavy chain variable region is shown as SEQ ID NO. 3. The anti-human CD15 engineering antibody can be used for diagnosing Hodgkin's lymphoma, leukemia immunophenotyping and differential diagnosis of mesothelioma and adenoma.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to an anti-human CD15 engineering antibody and application thereof.
Background
At present, the treatment of the Hodgkin lymphoma mainly adopts a combined treatment scheme taking chemotherapy as a main treatment and radiotherapy as an auxiliary treatment, and adopts chemotherapeutic drugs for controlling proliferation and inducing apoptosis. The medicines have serious damage to normal hematopoietic cells while killing leukemia cells, so that the medicines have serious toxic and side effects on organisms.
Hodgkin's Lymphoma (HL) is a unique type of lymphoma, accounting for about 10-20% of all lymphomas. Including two types of diseases: nodular lymphocytes are the master HL (NLPHL) and Classical HL (CHL). These two types of HL share some common features: there are only a few neoplastic large cells in diseased tissue-hodgkin cells and Reed-Sternberg (R-S) cells, with a large number of reactive non-neoplastic cells surrounding the neoplastic cells.
Radiotherapy has delayed toxicity to normal tissues. As survival time increases, the proportion of serious complications increases. In 20-30% of long-term survivors, secondary tumor and cardiotoxicity due to radiotherapy can occur. These long-term toxicities not only affect the quality of life of the patient, but also are important reasons for reducing the long-term survival rate of patients with good early prognosis. Therefore, there is a need to try to treat patients with good early prognosis using pure chemotherapy, and some related control studies have been carried out to compare pure chemotherapy with chemo-radiotherapy combination therapies such as EORTC, GELAH9F, CCG, tata, NCIC, MSKCC, etc., and the study results are not completely consistent, but most of the studies indicate that pure chemotherapy causes slight decrease in EFS or PFS compared to the combination therapy, without significant effect on 0S.
With the development of HL pathology, biology and immunology, some targeted drugs are also undergoing preclinical and clinical trials. SGNd5 is a compound of CD30 mab acting against synthetic anti-microtubules. The antibody-drug conjugate is stable in extracellular fluid, activates an anti-mitotic mechanism once entering tumor cells, can treat two types of lymphoma patients expressing CD30 antigen in a targeting manner, and shows better curative effect in vitro experiments. CD15 is taken as an antibody drug target and is expected to be applied to the treatment of Hodgkin lymphoma in later research.
Disclosure of Invention
In view of the above, the present invention aims to overcome the defects in the prior art and provides an anti-human CD15 engineering antibody and application thereof.
In order to achieve the above purpose, the technical scheme of the invention is realized as follows:
the invention provides an anti-human CD15 engineering antibody, which comprises a heavy chain variable region and a light chain variable region;
the complementarity determining regions of the heavy chain variable region include CDR1, CDR2 and CDR3; the amino acid sequence of CDR1 of the heavy chain variable region is shown as SEQ ID NO. 1; the amino acid sequence of CDR2 of the heavy chain variable region is shown as SEQ ID NO. 2; the amino acid sequence of CDR3 of the heavy chain variable region is shown as SEQ ID NO. 3;
the complementarity determining regions of the light chain variable region include CDR1, CDR2 and CDR3; the amino acid sequence of CDR1 of the light chain variable region is shown as SEQ ID NO. 4; the amino acid sequence of CDR2 of the light chain variable region is shown as SEQ ID NO. 5; the amino acid sequence of CDR3 of the light chain variable region is shown as SEQ ID NO. 6.
Further, the framework regions of the heavy chain variable region include FR1, FR2, FR3 and FR4; the amino acid sequence of FR1 of the heavy chain variable region is shown as SEQ ID NO. 7; the amino acid sequence of FR2 of the heavy chain variable region is shown as SEQ ID NO. 8; the amino acid sequence of FR3 of the heavy chain variable region is shown as SEQ ID NO. 9; the amino acid sequence of FR4 of the heavy chain variable region is shown as SEQ ID NO. 10;
the framework regions of the light chain variable region comprise FR1, FR2, FR3 and FR4; the amino acid sequence of FR1 of the light chain variable region is shown as SEQ ID NO. 11; the amino acid sequence of FR2 of the light chain variable region is shown as SEQ ID NO. 12; the amino acid sequence of FR3 of the light chain variable region is shown as SEQ ID NO. 13; the amino acid sequence of FR4 of the light chain variable region is shown as SEQ ID NO. 14.
Further, the amino acid sequence of the heavy chain variable region of the engineering antibody is shown as SEQ ID NO.15, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 16.
Further, the anti-human CD15 engineered antibody is of the IgG1 subtype.
The invention also provides a nucleotide molecule which codes for the anti-human CD15 engineering antibody.
Further, the nucleotide sequence of the nucleotide molecule coding the heavy chain variable region of the anti-human CD15 engineering antibody is shown as SEQ ID NO. 17, and the nucleotide sequence of the coding light chain variable region is shown as SEQ ID NO. 18.
The invention also provides an expression vector, which comprises the nucleotide molecule. The expression vector is pcDNA3.4 or Simple-T.
The invention also provides application of the anti-human CD15 engineering antibody, and application of the anti-human CD15 engineering antibody in preparation of a reagent for tumor diagnosis; the tumor is one of Hodgkin lymphoma, mesothelioma, adenoma or leukemia; the Hodgkin's lymphoma is classical Hodgkin's lymphoma; the adenoma is kidney-derived adenoma and colorectal adenocarcinoma; the application of the anti-human CD15 engineering antibody in preparing a reagent for the immunophenotyping of leukemia.
The invention also provides application of the expression vector in preparation of a reagent for tumor diagnosis; the tumor is one of Hodgkin lymphoma, mesothelioma, adenoma or leukemia; the Hodgkin's lymphoma is classical Hodgkin's lymphoma; the adenoma is kidney-derived adenoma and colorectal adenocarcinoma; the application of the expression vector in preparing a reagent for the immunophenotyping of leukemia.
The invention also provides an immunoassay kit which contains the anti-human CD15 engineering antibody and/or the expression vector.
Compared with the prior art, the invention has the following advantages:
the anti-human CD15 engineering antibody can be used for diagnosing Hodgkin's lymphoma, leukemia immunophenotyping and differential diagnosis of mesothelioma and adenoma.
The anti-human CD15 engineering antibody can specifically bind to human CD15 molecules, and the CD15 antibody subtype is changed from IgM to IgG1 subtype, thereby being beneficial to the purification, marking and detection of the antibody in the later period.
Drawings
FIG. 1 is an electrophoretogram of the purified product of example 3 of the present invention: non-reducing electrophoresis is arranged on the left side, and reducing electrophoresis is arranged on the right side;
FIG. 2 is a graph showing the activity of the purified product according to example 4 of the present invention: wherein, A is a control, B is an addition amount of 1 mug, and C is an addition amount of 0.5 mug;
FIG. 3 is a graph showing the activity detection of CD15-APC according to example 6 of the present invention: wherein A is control and lymphocyte, B is 0.25 μ gCD15-APC and lymphocyte, and C is 0.06 μ gCD15-APC and lymphocyte.
Detailed Description
Unless defined otherwise, technical terms used in the following examples have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains. The test reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods are conventional methods unless otherwise specified.
The present invention will be described in detail with reference to examples.
EXAMPLE 1 cloning of the light and heavy chain variable region Gene of mouse anti-human CD15 antibody
1. RNA extraction
The Trizol one-step method was used: (1) Taking hybridoma cells about 1×10 6 Adding 1ml of Trizol, blowing and mixing uniformly, and standing at room temperature for 10 minutes; (2) Adding 0.2ml of chloroform, shaking vigorously for 15 seconds, and standing at room temperature for 2-3 minutes; (3) 12000rpm of the powder at the time of the powder,centrifuging for 15 minutes at 4 ℃; (4) Taking the supernatant, adding 0.5ml of isopropanol, and standing for 15 minutes at room temperature; (5) 12000rpm,4 ℃, centrifugation for 15 minutes; (6) The supernatant was discarded, 1ml of 75% ethanol was added for washing, and the mixture was centrifuged at 7500rpm at 4℃for 5 minutes; (7) Discarding the supernatant, air-drying the precipitate, and adding 30 mu L of DEPC water for dissolution;
2. reverse transcription into cDNA (40 μl)
Reverse transcription reaction is carried out by using a TransScript All-in-one First-Strand cDNA Synthesis SuperMix for qPCR kit, 1.0 mug of total RNA is taken, 4 mug of TransScript All-in-one SuperMix for qPCR mug, 4 mug of gDNA remover, and DEPC water are supplemented to 20 mug, and after reaction is carried out at 42 ℃ for 15min, incubation is carried out at 85 ℃ for 5min, and the obtained product is preserved at 20 ℃;
3. PCR amplification of the light and heavy chain variable region genes of CD15 antibodies
Light chain variable region gene PCR amplification reaction System (50. Mu.l): designing universal degenerate primers: the upstream primer 5'-GACATT GTG CTC ACC CAG WCT SMH-3' (SEQ ID NO: 19), the downstream primer 5'-CCG TTAGAT CTC CAR BTT KGT SCS-3' (SEQ ID NO: 20); using cDNA as a template, and amplifying the pfu DNA polymerase with high fidelity; the PCR cycle was carried out at 94℃for 5 minutes; 94℃for 30 seconds, 55℃for 30 seconds, 72℃for 30 seconds, 30 cycles total; finally, the mixture is extended for 10 minutes at 72 ℃;
heavy chain variable region gene PCR amplification reaction System (50. Mu.l): an upstream primer 5'-CAG GTS MARCTG CAGSAG TCW GG-3' (SEQ ID NO: 21); the downstream primer 5'-TGA GGA GAC KGT GAC HGT GGT SCC-3' (SEQ ID NO: 22); using cDNA as a template, and amplifying the pfu DNA polymerase with high fidelity; the PCR cycle was carried out at 94℃for 5 minutes; 94 ℃,30 seconds, 55 ℃,30 seconds, 72 ℃,30 seconds, 35 cycles total; finally, the mixture is extended for 10 minutes at 72 ℃;
4. construction of sequencing vectors
pClone007 Blunt Simple Vector Kit is purchased from a family of organisms; the PCR product of the light and heavy chain variable region gene is recovered, is connected with a pClone007 Blunt Simple Vector carrier, is transformed by calcium chloride according to a conventional method, is screened for positive clones at an ampicillin concentration of 100 mu g/ml, is sent to a sequencing system, and completely accords with the characteristics of a plurality of conserved framework amino acids of an antibody in a protein database, and the sequence is an antibody gene sequence. Named pClone007-VH and pClone007-VL, respectively.
Example 2 construction of engineering antibody expression vectors pcDNA3.4-H and pcDNA3.4-L
Primers for amplifying heavy chain variable region (PDH-F and PDH-R), primers for amplifying light chain variable region (PDL-F and PDL-R), primers for amplifying heavy chain vector (ZTH-F and ZTH-R) and primers for amplifying light chain vector (ZTL-F and ZTL-R) were designed and synthesized based on the sequences of light and heavy chain variable region sequences and the sequence of vector pcDNA3.4 containing mouse light and heavy chain constant region.
PDH-F：
5’-TTCCAGGTTCCACTGGTGACGAGGTGAAGCTGCTCGAGTCC-3’（SEQ ID NO:23）；
PDH-R：
5’-GATGGGGGTGTCGTTTTGGCTGAGGAGACGGTGAGTGTGGTC-3’（SEQ ID NO:24） ；
PDL-F：
5’-TTCCAGGTTCCACTGGTGACGACATCGTCATGACTCAGGCTG-3’ （SEQ ID NO:25） ；
PDL-R：
5’-ACAGTTGGTGCAGCATCAGCCCTCTTGATCTCGAGCTTTGTTC-3’（SEQ ID NO:26） ；
ZTH-F：
5’-GCCAAAACGACACCCCCA-3’（SEQ ID NO:27）；
ZTH-R：
5’-GTCACCAGTGGAACCTGGAACC-3’（SEQ ID NO:28）；
ZTL-F：
5’-GCTGATGCTGCACCAACTGTAT-3’（SEQ ID NO:29）；
ZTL-R：
5’-GTCACCAGTGGAACCTGGAACC-3’（SEQ ID NO:30）；
Light chain variable region gene PCR amplification reaction System (50. Mu.l): an upstream primer PDL-F; a downstream primer PDL-R; using pClone007-VL as a template, and amplifying the pfu DNA polymerase with high fidelity; the PCR cycle was carried out at 94℃for 5 minutes; 94℃for 30 seconds, 58℃for 60 seconds, 72℃for 30 seconds, 30 cycles total; finally, the extension is carried out at 72 ℃ for 10 minutes.
Heavy chain variable region gene PCR amplification reaction System (50. Mu.l): an upstream primer PDH-F; a downstream primer PDH-R; using pClone007-VH as a template, and amplifying the pfu DNA polymerase with high fidelity; the PCR cycle was carried out at 94℃for 5 minutes; 94℃for 30 seconds, 58℃for 60 seconds, 72℃for 30 seconds, 30 cycles total; finally, the extension is carried out at 72 ℃ for 10 minutes.
Amplification of light chain linearization vector: an upstream primer ZTL-F; a downstream primer ZTL-R; using pcDNA3.4-CD34-L (synthetic sequence, containing mouse light chain constant region nucleotide sequence) as template, amplifying with high-fidelity pfu DNA polymerase; the PCR cycle was carried out at 94℃for 5 minutes; 94℃for 30 seconds, 58℃for 60 seconds, 72℃for 4 minutes, 30 cycles in total; finally, the extension is carried out at 72 ℃ for 10 minutes.
Amplification of heavy chain linearization vector: an upstream primer ZTH-F; a downstream primer ZTH-R; using pcDNA-CD34-H (synthetic sequence, including mouse heavy chain constant region nucleotide sequence) as template, high-fidelity pfu DNA polymerase amplification; the PCR cycle was carried out at 94℃for 5 minutes; 94℃for 30 seconds, 58℃for 60 seconds, 72℃for 4 minutes, 30 cycles in total; finally, the extension is carried out at 72 ℃ for 10 minutes.
And respectively recovering PCR products, respectively recombining the recovered light chain variable region PCR products and heavy chain variable region PCR products with the recovered linearization carrier products by using recombinase, converting DH5 alpha clone strains after recombination, picking single bacterial colony for amplification culture, carrying out sample feeding and sequencing, and using clones with correct sequencing for transfection of 293 cells.
EXAMPLE 3 expression and purification of an anti-human CD15 engineering antibody
1. Expression of engineered antibody CD15 antibodies
Taking out the prepared 293 cells from the incubator, preparing two 15ml sterile centrifuge tubes, adding 5ml SPM culture medium and 100 mug sterile plasmid DNA (containing light chain expression vector and heavy chain expression vector) into one of the two sterile centrifuge tubes, and gently beating and uniformly mixing; taking the other separation tube, adding 5ml of SPM and 500 μl of transfection reagent, lightly blowing and mixing; transferring all liquid in the centrifuge tube containing the transfection reagent into the centrifuge tube containing the plasmid, and lightly blowing and uniformly mixing; standing for 10 minutes at room temperature to prepare a plasmid-carrier compound; taking out the fine powder from the constant temperature shaking tableAdding the prepared plasmid-vector complex while shaking the cells, and returning CO 2 Shake culturing in a constant temperature shaking table; supplementing auxiliary materials on the 1 st day, and harvesting cell supernatants on the 6 th day;
2. purification of anti-human CD15 engineered antibodies
Purifying the cell supernatant collected in the previous step by using a Protein A affinity column, balancing the column by using PBS, loading the sample, eluting by using glycine with the pH value of 3.0, replacing the solution by using a G25 column for eluting pure antibody, replacing the solution by using PBS, sampling for electrophoresis test, sub-packaging, and freezing at the temperature of-20 ℃, wherein the electrophoresis result is shown in figure 1.
3. Western blot identification: western blotting was performed with the main reference molecular cloning method, and the results demonstrated that the bands were antibodies.
EXAMPLE 4 Activity assay of anti-human CD15 engineering antibodies
Detecting peripheral blood of normal people: 100 mu l of normal anticoagulated peripheral blood is added into each tube, different amounts of engineering antibodies CD15 (1 mu g and 0.5 mu g) are respectively added, and the mixture is incubated for 30 minutes at room temperature and in a dark place; adding 2ml of hemolysin, reacting at room temperature in a dark place for 10 minutes, centrifuging at 1000 rpm for 5 minutes, and discarding the supernatant; adding 2ml of cold PBS buffer, re-suspending, centrifuging at 1000 rpm for 5 minutes, and discarding the supernatant; then 0.5 mug of APC marked mouse secondary antibody is added, and incubated for 20 minutes at room temperature and in dark place; adding 2ml of cold PBS buffer, re-suspending, centrifuging at 1000 rpm for 5 minutes, and discarding the supernatant; adding the matched antibodies CD45-PerCP and CD33-FITC, reacting for 20 minutes, adding 2ml of cold PBS buffer solution, re-suspending, centrifuging for 5 minutes at 1000 revolutions per minute, and discarding the supernatant; 250 μl PBS buffer was added and flow cytometer detected.
As shown in FIG. 2, the CD15 pure antibody reacted normally on granulocytes, and the specificity was not a problem.
EXAMPLE 5 APC labelling of anti-human CD15 engineered antibodies
Reducing the anti-human CD15 engineering antibody by using a reducing agent, uniformly mixing the reducing agent with the activated APC, stirring at 25 ℃, and reacting for 1 hour; and purifying by using an S200 increment purification column to obtain the APC marked CD15 engineering antibody.
EXAMPLE 6 Activity assay of anti-human CD15-APC engineering antibody
Detecting peripheral blood of normal people: 100 mu l of normal anticoagulated peripheral blood is added into each tube, different amounts of engineering antibodies CD15-APC (0.25 mu g and 0.06 mu g) are respectively added, and then antibodies CD45-PercP and CD33-FITC are added for incubation for 30 minutes at room temperature and in a dark place; adding 2ml of hemolysin, reacting at room temperature in a dark place for 10 minutes, centrifuging at 1000 rpm for 5 minutes, and discarding the supernatant; adding 2ml of cold PBS buffer, centrifuging at 1000 rpm for 5 minutes, and discarding the supernatant; 250 μl PBS buffer was added and flow cytometer detected.
As shown in FIG. 3, CD15-APC reacts normally on granulocytes, the specificity is not a problem, and the activity of the labeled CD15 antibody is not affected.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (10)
1. An anti-human CD15 engineered antibody, characterized in that: the engineered antibody comprises a heavy chain variable region and a light chain variable region;
the complementarity determining regions of the heavy chain variable region include CDR1, CDR2 and CDR3; the amino acid sequence of CDR1 of the heavy chain variable region is shown as SEQ ID NO. 1; the amino acid sequence of CDR2 of the heavy chain variable region is shown as SEQ ID NO. 2; the amino acid sequence of CDR3 of the heavy chain variable region is shown as SEQ ID NO. 3;
the complementarity determining regions of the light chain variable region include CDR1, CDR2 and CDR3; the amino acid sequence of CDR1 of the light chain variable region is shown as SEQ ID NO. 4; the amino acid sequence of CDR2 of the light chain variable region is shown as SEQ ID NO. 5; the amino acid sequence of CDR3 of the light chain variable region is shown as SEQ ID NO. 6.
2. The anti-human CD15 engineered antibody of claim 1, wherein: the framework regions of the heavy chain variable region comprise FR1, FR2, FR3 and FR4; the amino acid sequence of FR1 of the heavy chain variable region is shown as SEQ ID NO. 7; the amino acid sequence of FR2 of the heavy chain variable region is shown as SEQ ID NO. 8; the amino acid sequence of FR3 of the heavy chain variable region is shown as SEQ ID NO. 9; the amino acid sequence of FR4 of the heavy chain variable region is shown as SEQ ID NO. 10;
the framework regions of the light chain variable region comprise FR1, FR2, FR3 and FR4; the amino acid sequence of FR1 of the light chain variable region is shown as SEQ ID NO. 11; the amino acid sequence of FR2 of the light chain variable region is shown as SEQ ID NO. 12; the amino acid sequence of FR3 of the light chain variable region is shown as SEQ ID NO. 13; the amino acid sequence of FR4 of the light chain variable region is shown as SEQ ID NO. 14.
3. The anti-human CD15 engineered antibody of claim 1, wherein: the amino acid sequence of the heavy chain variable region of the engineering antibody is shown as SEQ ID NO.15, and the amino acid sequence of the light chain variable region is shown as SEQ ID NO. 16.
4. The anti-human CD15 engineered antibody of claim 1, wherein: the anti-human CD15 engineering antibody is of an IgG1 subtype.
5. A nucleic acid molecule characterized in that: the nucleic acid molecule encodes the anti-human CD15 engineered antibody of any one of claims 1-4.
6. The nucleic acid molecule of claim 5, wherein: the nucleotide sequence of the nucleic acid molecule encoding the heavy chain variable region of the anti-human CD15 engineering antibody is shown as SEQ ID NO. 17, and the nucleotide sequence encoding the light chain variable region is shown as SEQ ID NO. 18.
7. An expression vector, characterized in that: the expression vector comprises the nucleic acid molecule of claim 5 or 6.
8. Use of an anti-human CD15 engineered antibody according to any one of claims 1-4, characterized in that: the application of the anti-human CD15 engineering antibody in preparing a reagent for tumor diagnosis or in preparing a reagent for immunophenotyping of leukemia; the tumor is Hodgkin lymphoma; the Hodgkin's lymphoma is classical Hodgkin's lymphoma.
9. Use of the expression vector of claim 7, wherein: the application of the expression vector in preparing a reagent for tumor diagnosis or in preparing a reagent for immunophenotyping of leukemia; the tumor is Hodgkin lymphoma; the Hodgkin's lymphoma is classical Hodgkin's lymphoma.
10. An immunoassay kit, characterized in that: the kit contains the anti-human CD15 engineering antibody according to any one of claims 1-4 and/or the expression vector according to claim 7.
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| CN101885770A (en) * | 2009-05-12 | 2010-11-17 | 中国医药集团总公司四川抗菌素工业研究所 | Fully human antibody capable of specifically bonding CD15 and use thereof |
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| CN101885770A (en) * | 2009-05-12 | 2010-11-17 | 中国医药集团总公司四川抗菌素工业研究所 | Fully human antibody capable of specifically bonding CD15 and use thereof |
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| Title |
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| Wilma Pellegrini等.MMA monoclonal antibody is a superior anti-CD15 reagent for the diagnosis of classical Hodgkin's lymphoma?.Haematologica.2007,第92卷(第5期),第708-709页. * |
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