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CN108484772A - The humanized antibody H5L5 of anti-HER2 antigens and its application - Google Patents

️Tue Sep 04 2018

CN108484772A - The humanized antibody H5L5 of anti-HER2 antigens and its application - Google Patents

The humanized antibody H5L5 of anti-HER2 antigens and its application Download PDF

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Publication number

CN108484772A

CN108484772A CN201810320106.8A CN201810320106A CN108484772A CN 108484772 A CN108484772 A CN 108484772A CN 201810320106 A CN201810320106 A CN 201810320106A CN 108484772 A CN108484772 A CN 108484772A Authority

China

Prior art keywords

sequence

ser

amino acids

val

thr

Prior art date

2018-04-11

Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)

Pending

Application number

CN201810320106.8A

Other languages

Chinese (zh)

Inventor

曹诚

周瑞

朱林

靳彦文

张部昌

Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)

Institute of Pharmacology and Toxicology of AMMS

Anhui University

Original Assignee

Institute of Pharmacology and Toxicology of AMMS

Anhui University

Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)

2018-04-11

Filing date

2018-04-11

Publication date

2018-09-04

2018-04-11 Application filed by Institute of Pharmacology and Toxicology of AMMS, Anhui University filed Critical Institute of Pharmacology and Toxicology of AMMS

2018-04-11 Priority to CN201810320106.8A priority Critical patent/CN108484772A/en

2018-09-04 Publication of CN108484772A publication Critical patent/CN108484772A/en

Status Pending legal-status Critical Current

Links

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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 kind of humanized antibody H5L5 of anti-HER2 antigens and its applications.The present invention provides a kind of IgG, CDR1, CDR2 and CDR3 in its heavy chain variable region are successively if the sequence of sequence table 1 is from shown in the 50th 54 amino acids residue of N-terminal, the 69th 85 amino acids residue and the 118th 128 amino acids residue, and CDR1, CDR2 and CDR3 in light chain variable region are successively if the sequence of sequence table 3 is from shown in the 44th 54 amino acids residue of N-terminal, the 70th 76 amino acids residue and the 109th 117 amino acids residue.Experimental result confirms that antibody provided by the invention has good combination activity with HER2 antigens and promotes the activity of apoptosis of tumor cells, and has different epitopes from Herceptin.Humanized antibody H5L5 provided by the invention and preparation method thereof will have broad application prospects in the field for the treatment of tumour.

Description

抗HER2抗原的人源化抗体H5L5及其应用Humanized antibody H5L5 against HER2 antigen and its application

技术领域technical field

本发明涉及一种抗HER2抗原的人源化抗体H5L5及其应用。The invention relates to an anti-HER2 antigen humanized antibody H5L5 and its application.

背景技术Background technique

乳腺癌是一种严重危害女性健康的恶性肿瘤，全球每年新增乳腺癌约有120万人，发病率占全身各种恶性肿瘤的7-10％。乳腺癌的治疗方式目前主要包括手术治疗、化疗、放疗与生物治疗。1987年，研究发现在乳腺癌亚型中大约有15-20％乳腺癌细胞表面过量表达人表皮生长因子受体2(HER2)。HER2蛋白是具有受体酪氨酸激酶活性的跨膜糖蛋白，属表皮生长因子受体家族(包括EGFR、HER2、HER3与HER4)成员之一。HER2在细胞分化与增殖中发挥重要作用，HER2过量表达的乳腺癌一般预后不良，如生存率低、复发间隔短与转移发生率增加。因此，HER2可以作为乳腺癌生物靶向治疗的重要靶标。Breast cancer is a malignant tumor that seriously endangers women's health. There are about 1.2 million new breast cancers in the world every year, and the incidence rate accounts for 7-10% of all kinds of malignant tumors in the whole body. Breast cancer treatment methods currently mainly include surgery, chemotherapy, radiotherapy and biological therapy. In 1987, it was found that about 15-20% of breast cancer subtypes overexpress human epidermal growth factor receptor 2 (HER2) on the surface of breast cancer cells. HER2 protein is a transmembrane glycoprotein with receptor tyrosine kinase activity, which is a member of the epidermal growth factor receptor family (including EGFR, HER2, HER3 and HER4). HER2 plays an important role in cell differentiation and proliferation. Breast cancer with overexpression of HER2 generally has a poor prognosis, such as low survival rate, short recurrence interval and increased incidence of metastasis. Therefore, HER2 can be used as an important target for biological targeted therapy of breast cancer.

1998年，美国FDA批准靶向HER2人源化单克隆抗体——曲妥珠单抗(trastuzumab，又名赫赛汀)用于HER2阳性乳腺癌治疗的一线药物。2015年，曲妥珠单抗的全球销售额达65.6亿美元。曲妥珠单抗对HER2信号通路的影响包括细胞周期阻滞、细胞凋亡与血管生成抑制等。目前关于曲妥珠单抗的作用机制主要包括：PI3K/AKT和MAPK信号通路的抑制作用；阻止HER2胞外区的裂解；抗体依赖的细胞介导的细胞毒作用和补体依赖的细胞毒作用。In 1998, the US FDA approved the humanized monoclonal antibody targeting HER2—trastuzumab (also known as Herceptin) as a first-line drug for the treatment of HER2-positive breast cancer. In 2015, the global sales of trastuzumab reached 6.56 billion US dollars. The effects of trastuzumab on the HER2 signaling pathway include cell cycle arrest, apoptosis and angiogenesis inhibition. The current mechanism of action of trastuzumab mainly includes: inhibition of PI3K/AKT and MAPK signaling pathways; prevention of cleavage of the extracellular region of HER2; antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity.

曲妥珠单抗可显著提高HER2高表达乳腺癌的治疗效果，然而，曲妥珠单抗的原发性和获得性耐受给临床带来较大的挑战，因此研发出亲和力高或具有不同靶标位置的人源化抗体显得尤为重要。Trastuzumab can significantly improve the therapeutic effect of breast cancer with high HER2 expression. However, the primary and acquired resistance of trastuzumab has brought great challenges to the clinic. Humanized antibodies to target sites are particularly important.

发明内容Contents of the invention

本发明的目的是提供一种抗HER2抗原的人源化抗体H5L5及其应用。The purpose of the present invention is to provide an anti-HER2 antigen humanized antibody H5L5 and its application.

本发明提供了一种IgG(命名为H5L5抗体)，其重链可变区中的CDR1、CDR2和CDR3依次如序列表的序列1自N端第50-54位氨基酸残基、第69-85位氨基酸残基和第118-128位氨基酸残基所示，其轻链可变区中的CDR1、CDR2和CDR3依次如序列表的序列3自N端第44-54位氨基酸残基、第70-76位氨基酸残基和第109-117位氨基酸残基所示。The present invention provides an IgG (named as H5L5 antibody), the CDR1, CDR2 and CDR3 in the variable region of the heavy chain are sequentially as shown in sequence 1 of the sequence listing from the 50th to 54th amino acid residues at the N-terminal, and the 69th to 85th amino acid residues. amino acid residues and 118-128 amino acid residues, CDR1, CDR2 and CDR3 in the light chain variable region are sequentially as shown in sequence 3 of the sequence listing from N-terminal 44-54 amino acid residues, 70 -76 amino acid residues and 109-117 amino acid residues.

所述重链可变区如序列表的序列1自N端第20-139位氨基酸残基所示。The heavy chain variable region is shown in the 20th-139th amino acid residues from the N-terminal of Sequence 1 in the sequence listing.

所述轻链可变区如序列表的序列3自N端第21-128位氨基酸残基所示。The light chain variable region is shown in the 21st-128th amino acid residues from the N-terminal of the sequence 3 in the sequence listing.

所述重链为如下(a1)或(a2)：(a1)序列表的序列1自N端第20-469位氨基酸残基组成的蛋白质；(a2)序列表的序列1所示的蛋白质。The heavy chain is as follows (a1) or (a2): (a1) a protein composed of amino acid residues from N-terminal 20-469 in sequence 1 of the sequence listing; (a2) a protein shown in sequence 1 of the sequence listing.

所述轻链为如下(b1)或(b2)：(b1)序列表的序列3自N端第21-234位氨基酸残基组成的蛋白质；(b2)序列表的序列3所示的蛋白质。The light chain is the following (b1) or (b2): (b1) a protein composed of amino acid residues 21-234 at the N-terminus of sequence 3 in the sequence listing; (b2) a protein shown in sequence 3 in the sequence listing.

编码以上任一所述IgG的基因也属于本发明的保护范围。Any gene encoding IgG described above also falls within the protection scope of the present invention.

编码所述重链的基因为如下(c1)或(c2)或(c3)：The gene encoding the heavy chain is as follows (c1) or (c2) or (c3):

(c1)序列表的序列2自5’端第77-1426位核苷酸所示的DNA分子；(c1) the DNA molecule shown in the 77th-1426th nucleotide of the sequence 2 of the sequence listing from the 5' end;

(c2)序列表的序列2自5’端第20-1429位核苷酸所示的DNA分子；(c2) the DNA molecule shown in the 20th-1429th nucleotide from the 5' end of sequence 2 of the sequence listing;

(c3)序列表的序列2所示的DNA分子。(c3) A DNA molecule shown in Sequence 2 of the Sequence Listing.

编码所述轻链的基因为如下(d1)或(d2)或(d3)：The gene encoding said light chain is as follows (d1) or (d2) or (d3):

(d1)序列表的序列4自5’端第80-821位核苷酸所示的DNA分子；(d1) the DNA molecule shown in the 80th-821st nucleotide from the 5' end of sequence 4 of the sequence listing;

(d2)序列表的序列4自5’端第20-724位核苷酸所示的DNA分子；(d2) the DNA molecule shown in the 20th-724th nucleotide from the 5' end of sequence 4 of the sequence listing;

(d3)序列表的序列4所示的DNA分子。(d3) DNA molecule shown in sequence 4 of the sequence listing.

本发明还保护以上任一所述IgG在制备用于诱导肿瘤细胞凋亡的药物中的应用。The present invention also protects the use of any one of the above IgGs in the preparation of drugs for inducing tumor cell apoptosis.

本发明还保护以上任一所述IgG在制备用于治疗肿瘤的药物中的应用。The present invention also protects the use of any one of the above IgGs in the preparation of drugs for treating tumors.

本发明还保护以上任一所述IgG在制备产品中的应用；所述产品的用途为如下(f1)或(f2)或(f3)或(f4)：The present invention also protects the application of any of the above IgGs in the preparation of products; the use of the products is as follows (f1) or (f2) or (f3) or (f4):

(f1)检测肿瘤细胞；(f1) detecting tumor cells;

(f2)结合肿瘤细胞；(f2) binding to tumor cells;

(f3)检测人表皮生长因子受体2；(f3) detecting human epidermal growth factor receptor 2;

(f4)结合人表皮生长因子受体2。(f4) Binds human epidermal growth factor receptor 2.

本发明还保护一种产品，其活性成分为任一所述IgG；所述产品的用途为如下(f1)或(f2)或(f3)或(f4)：The present invention also protects a product whose active ingredient is any one of the IgGs; the use of the product is as follows (f1) or (f2) or (f3) or (f4):

(f1)检测肿瘤细胞；(f1) detecting tumor cells;

(f2)结合肿瘤细胞；(f2) binding to tumor cells;

(f3)检测人表皮生长因子受体2；(f3) detecting human epidermal growth factor receptor 2;

(f4)结合人表皮生长因子受体2。(f4) Binds human epidermal growth factor receptor 2.

以上任一所述肿瘤细胞可为高表达人表皮生长因子受体2的肿瘤细胞。所述肿瘤细胞具体可为乳腺癌细胞，例如SK-BR-3细胞。Any of the above tumor cells may be tumor cells that highly express human epidermal growth factor receptor 2. The tumor cells can specifically be breast cancer cells, such as SK-BR-3 cells.

以上任一所述肿瘤为可为由高表达人表皮生长因子受体2的肿瘤细胞引起的肿瘤。所述肿瘤具体可为乳腺癌。Any of the tumors mentioned above may be tumors caused by tumor cells that highly express human epidermal growth factor receptor 2. The tumor may specifically be breast cancer.

本发明的发明人制备了一种抗HER2抗原的人源化抗体H5L5。将抗体重链序列与轻链序列克隆、表达与纯化后，通过亲和力分析筛选到亲和力高且具有不同抗原表位的人源化抗体。The inventors of the present invention prepared a humanized antibody H5L5 against HER2 antigen. After cloning, expressing and purifying the antibody heavy chain sequence and light chain sequence, humanized antibodies with high affinity and different epitopes were screened by affinity analysis.

实验结果证实，本发明提供的人源化抗体H5L5与HER2抗原具有良好的结合活性和促使肿瘤细胞凋亡的活性。本发明的人源化抗体能更好与HER2结合，且与曲妥珠单抗具有不同的抗原表位，通过与曲妥珠单抗联合使用或构建双特异性抗体进而为治疗乳腺癌提供新的思路。本发明提供的人源化抗体H5L5及其制备方法在治疗肿瘤的领域将有广阔的应用前景。Experimental results confirm that the humanized antibody H5L5 provided by the present invention has good binding activity to HER2 antigen and the activity of promoting tumor cell apoptosis. The humanized antibody of the present invention can better bind to HER2, and has a different antigenic epitope from trastuzumab, and provides a new treatment for breast cancer by using it in combination with trastuzumab or constructing a bispecific antibody. train of thought. The humanized antibody H5L5 provided by the invention and the preparation method thereof will have broad application prospects in the field of treating tumors.

附图说明Description of drawings

图1为洗脱曲线。Figure 1 is the elution profile.

图2为抗体电泳检测结果。Figure 2 shows the results of antibody electrophoresis detection.

图3为ELISA检测结果。Figure 3 is the result of ELISA detection.

图4为流式细胞术检测结果。Figure 4 shows the results of flow cytometry detection.

图5为细胞凋亡检测结果。Figure 5 shows the results of cell apoptosis detection.

图6为竞争性抑制实验检测结果。Figure 6 is the test result of competitive inhibition experiment.

具体实施方式Detailed ways

以下的实施例便于更好地理解本发明，但并不限定本发明。下述实施例中的实验方法，如无特殊说明，均为常规方法。下述实施例中所用的试验材料，如无特殊说明，均为自常规生化试剂商店购买得到的。以下实施例中的定量试验，均设置三次重复实验，结果取平均值。The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent stores. Quantitative experiments in the following examples were all set up to repeat the experiments three times, and the results were averaged.

pcDNA-3.3载体(线性质粒)：Invitrogen公司，目录号：K8300-01。pcDNA-3.3 vector (linear plasmid): Invitrogen, catalog number: K8300-01.

pOptiVEC载体(线性质粒)：Invitrogen公司，目录号：12744-017。pOptiVEC vector (linear plasmid): Invitrogen, catalog number: 12744-017.

HEK293F细胞：普诺赛公司(Procell),货号：CL-0313。HEK293F cells: Procell, product number: CL-0313.

SK-BR-3细胞：上海斯信生物科技有限公司，货号：LOOKBIO SK-BR-3。SK-BR-3 cells: Shanghai Sixin Biotechnology Co., Ltd., product number: LOOKBIO SK-BR-3.

I培养基(Opti-MEMI Reduced Serum Medium)：Invitrogen公司，目录号：31985-062。 I medium (Opti-MEMI Reduced Serum Medium): Invitrogen Company, catalog number: 31985-062.

脂质体(293fectin^TMTransfection Reagent)：Invitrogen公司，目录号：12347-019。Liposome (293fectin ^™ Transfection Reagent): Invitrogen, catalog number: 12347-019.

FreeStyle^TM293表达培养基：Invitrogen公司，目录号：12338-018。FreeStyle ^™ 293 Expression Medium: Invitrogen, catalog number: 12338-018.

Human IgG：中杉金桥(ZSGB-BIO)，货号：ZDR-5001。Human IgG: Zhongshan Jinqiao (ZSGB-BIO), product number: ZDR-5001.

实施例1、抗体的发现Embodiment 1, the discovery of antibody

发明人经过大量摸索、分析、验证，设计了一种新的抗HER2抗原的抗体(命名为H5L5抗体)。H5L5抗体为IgG，其重链如序列表的序列1所示，其轻链如序列表的序列3所示。The inventor designed a new anti-HER2 antigen antibody (named H5L5 antibody) after a lot of exploration, analysis, and verification. The H5L5 antibody is IgG, its heavy chain is shown in sequence 1 of the sequence listing, and its light chain is shown in sequence 3 of the sequence listing.

序列表的序列1中，自N端第1-19位氨基酸残基组成前导肽，第20-139位氨基酸残基组成重链可变区VL(其中，第50-54位氨基酸残基组成CDR1，第69-85位氨基酸残基组成CDR2，第118-128位氨基酸残基组成CDR3)，第140-237位氨基酸残基组成重链恒定区CH1，第238-252位氨基酸残基组成重链铰链区Hinge，第253-363位氨基酸残基组成重链恒定区CH2，第364-469位氨基酸残基组成重链恒定区CH3。In Sequence 1 of the sequence listing, amino acid residues 1-19 from the N-terminal form the leader peptide, and amino acid residues 20-139 form the heavy chain variable region VL (among them, amino acid residues 50-54 form the CDR1 , amino acid residues 69-85 constitute CDR2, amino acid residues 118-128 constitute CDR3), amino acid residues 140-237 constitute the heavy chain constant region CH1, and amino acid residues 238-252 constitute the heavy chain In the hinge region Hinge, amino acid residues 253-363 constitute the heavy chain constant region CH2, and amino acid residues 364-469 constitute the heavy chain constant region CH3.

序列表的序列2所示的DNA分子编码序列表的序列1所示的多肽。序列表的序列2中，自5’端第20-76位核苷酸编码前导肽，第77-436位核苷酸编码VH，第437-730位核苷酸编码CH1，第731-775位核苷酸编码Hinge，第776-1108位核苷酸编码CH2，第1109-1426位核苷酸编码CH3，第1427-1429位核苷酸为终止密码子。The DNA molecule shown in Sequence 2 of the Sequence Listing encodes the polypeptide shown in Sequence 1 of the Sequence Listing. In sequence 2 of the sequence listing, nucleotides 20-76 from the 5' end encode a leader peptide, nucleotides 77-436 encode VH, nucleotides 437-730 encode CH1, and nucleotides 731-775 Nucleotides code for Hinge, nucleotides 776-1108 code for CH2, nucleotides 1109-1426 code for CH3, and nucleotides 1427-1429 are stop codons.

序列表的序列3中，自N端第1-20位氨基酸残基组成前导肽，第21-128位氨基酸残基组成轻链可变区VL(其中，第44-54位氨基酸残基组成CDR1，第70-76位氨基酸残基组成CDR2，第109-117位氨基酸残基组成CDR3)，第129-234位氨基酸残基组成轻链恒定区CL。In sequence 3 of the sequence listing, amino acid residues 1-20 from the N-terminal form the leader peptide, and amino acid residues 21-128 form the light chain variable region VL (among them, amino acid residues 44-54 form the CDR1 , amino acid residues 70-76 constitute CDR2, amino acid residues 109-117 constitute CDR3), and amino acid residues 129-234 constitute the light chain constant region CL.

序列表的序列4所示的DNA分子编码序列表的序列3所示的多肽。序列表的序列4中，自5’端第20-79位核苷酸编码前导肽，第80-403位核苷酸编码VL，第404-721位核苷酸编码CL，第722-724位核苷酸为终止密码子。The DNA molecule shown in sequence 4 of the sequence listing encodes the polypeptide shown in sequence 3 of the sequence listing. In sequence 4 of the sequence listing, nucleotides 20-79 from the 5' end encode a leader peptide, nucleotides 80-403 encode VL, nucleotides 404-721 encode CL, and nucleotides 722-724 Nucleotides are stop codons.

实施例2、抗体的制备Embodiment 2, the preparation of antibody

一、H5L5抗体的制备1. Preparation of H5L5 antibody

1、重组质粒的构建1. Construction of recombinant plasmids

(1)将序列表的序列2所示的DNA分子插入pOptiVEC载体，得到重组质粒pOptiVEC-H5。(1) Insert the DNA molecule shown in Sequence 2 of the Sequence Listing into the pOptiVEC vector to obtain the recombinant plasmid pOptiVEC-H5.

(2)将序列表的序列4所示的DNA分子插入pcDNA-3.3载体,得到重组质粒pcDNA-3.3-L5。(2) Insert the DNA molecule shown in sequence 4 in the sequence listing into the pcDNA-3.3 vector to obtain the recombinant plasmid pcDNA-3.3-L5.

2、抗体的制备2. Antibody preparation

(1)取15μg重组质粒pOptiVEC-H5和15μg重组质粒pcDNA3.3-L5，加入 I培养基定容至1ml，轻轻混匀。(1) Take 15 μg of recombinant plasmid pOptiVEC-H5 and 15 μg of recombinant plasmid pcDNA3.3-L5, add Dilute the volume of medium I to 1ml and mix gently.

(2)取60μl脂质体(293fectin^TMTransfection Reagent)，加入 I培养基定容至1ml，轻轻混匀后室温孵育5min。(2) Take 60μl liposome (293fectin ^TM Transfection Reagent), add Dilute the volume of medium I to 1ml, mix gently and incubate at room temperature for 5min.

(3)将步骤(1)的溶液和步骤(2)的溶液轻轻混匀，室温孵育20-30min。(3) Gently mix the solution of step (1) and step (2), and incubate at room temperature for 20-30 minutes.

(4)取125ml细胞培养瓶，加入28ml用Freestyle^TM 293表达培养基悬浮的293F细胞(细胞含量为3×10⁷)。(4) Take a 125ml cell culture flask and add 28ml of 293F cells suspended in Freestyle ^™ 293 expression medium (the cell content is 3×10 ⁷ ).

(5)将2ml步骤(3)的溶液逐滴加入至步骤(4)的细胞培养瓶中，在37℃、8％CO₂的生物培养床150rpm振荡培养至细胞活力低于50％(6-9天)。(5) the solution of 2ml step (3) is added dropwise in the cell culture bottle of step (4), at 37 ℃, 8% CO _150rpm shaking culture of biological culture bed until cell viability is lower than 50% (6- 9 days).

(6)完成步骤(5)后，12000rpm离心15min收集细胞培养上清，调pH至6.0-7.0后用0.45μm滤膜过滤，收集滤液。(6) After step (5), centrifuge at 12,000 rpm for 15 minutes to collect the cell culture supernatant, adjust the pH to 6.0-7.0, and filter with a 0.45 μm filter membrane to collect the filtrate.

(7)取步骤(6)得到的滤液，采用Bio-Rad Duo-Flow蛋白纯化系统(760-0135)进行纯化。(7) Take the filtrate obtained in step (6), and use Bio-Rad Duo-Flow protein purification system (760-0135) for purification.

Bio-Scale Mini Affi-prep proteinA亲和层析柱(伯乐公司，货号：732-4602)：柱体积为5毫升预装柱。Bio-Scale Mini Affi-prep proteinA affinity chromatography column (Bio-Rad, product number: 732-4602): the column volume is 5 ml prepacked column.

结合缓冲液(pH7.5)：含3M NaCl的20mM磷酸盐缓冲液。Binding buffer (pH7.5): 20 mM phosphate buffer containing 3M NaCl.

洗脱缓冲液(pH2.8)：0.1M柠檬酸盐缓冲液。Elution buffer (pH2.8): 0.1M citrate buffer.

流速：2ml/min。Flow rate: 2ml/min.

过程：(1)用50ml结合缓冲液平衡柱子；(2)上样；(3)用50ml结合缓冲液洗涤柱子；(4)用25ml洗脱缓冲液洗脱目的蛋白，收集保留UV280大于0.05AU的过柱后溶液，洗脱曲线见图1。Process: (1) equilibrate the column with 50ml binding buffer; (2) load the sample; (3) wash the column with 50ml binding buffer; (4) elute the target protein with 25ml elution buffer, collect and retain UV280 greater than 0.05AU The post-column solution, the elution curve is shown in Figure 1.

(8)取步骤(7)得到的过柱后溶液，立刻用Tris-Hcl溶液(pH9.0)调pH至7.0，然后使用截留分子量为10kD的超滤管(Millipore公司，Amicon Ultra-4)进行浓缩换液处理(浓缩换液采用的缓冲液为PBS缓冲液)，得到的溶液即为含H5L5抗体的溶液，命名为H5L5溶液。(8) Get the post-column solution obtained in step (7), adjust the pH to 7.0 with Tris-Hcl solution (pH9.0) immediately, and then use an ultrafiltration tube (Millipore company, Amicon Ultra-4) with a molecular weight cut-off of 10kD Concentrate and change the solution (the buffer used for the concentration and change is PBS buffer), and the obtained solution is the solution containing the H5L5 antibody, which is named as the H5L5 solution.

二、阳性对照抗体(Trastuzumab)的制备2. Preparation of positive control antibody (Trastuzumab)

采用曲妥珠单抗作为阳性对照。Trastuzumab was used as a positive control.

曲妥珠单抗，其重链可变区序列如序列表的序列5所示，其轻链可变区序列如序列表的序列6所示。序列表的序列7所示的DNA分子编码序列表的序列5所示的多肽。序列表的序列8所示的DNA分子编码序列表的序列6所示的多肽。Trastuzumab, its heavy chain variable region sequence is shown in sequence 5 in the sequence listing, and its light chain variable region sequence is shown in sequence 6 in the sequence listing. The DNA molecule shown in sequence 7 of the sequence listing encodes the polypeptide shown in sequence 5 of the sequence listing. The DNA molecule shown in sequence 8 of the sequence listing encodes the polypeptide shown in sequence 6 of the sequence listing.

参照步骤一制备得到阳性对照抗体溶液(Trastuzumab溶液)。Refer to Step 1 to prepare a positive control antibody solution (Trastuzumab solution).

抗体溶液的变性还原聚丙烯酰胺凝胶电泳见图2A，其中泳道1为阳性对照抗体溶液(Trastuzumab溶液)，泳道2为H5L5溶液，电泳可检测到抗体的重链与轻链。The denaturing reducing polyacrylamide gel electrophoresis of the antibody solution is shown in Figure 2A, where lane 1 is the positive control antibody solution (Trastuzumab solution), and lane 2 is the H5L5 solution. The heavy chain and light chain of the antibody can be detected by electrophoresis.

抗体溶液的变性非还原聚丙烯酰胺凝胶电泳见图2B，其中泳道1为阳性对照抗体溶液(Trastuzumab溶液)，泳道2为H5L5溶液，电泳可检测到抗体的大小。The denaturing non-reducing polyacrylamide gel electrophoresis of the antibody solution is shown in Figure 2B, wherein lane 1 is the positive control antibody solution (Trastuzumab solution), and lane 2 is the H5L5 solution, and the size of the antibody can be detected by electrophoresis.

三、空载体实验3. Empty vector experiment

用pOptiVEC载体代替重组质粒pOptiVEC-H5，用pcDNA-3.3载体代替重组质粒pcDNA-3.3-L5，按照步骤一的2进行操作，洗脱过程中不显示任何洗脱峰。Replace the recombinant plasmid pOptiVEC-H5 with the pOptiVEC vector, and replace the recombinant plasmid pcDNA-3.3-L5 with the pcDNA-3.3 vector, and operate according to step 2 of step 1, and no elution peak appears during the elution process.

实施例3、亲和力检测Embodiment 3, affinity detection

一、ELISA检测1. ELISA detection

1、包被1. Coating

取酶标板，每孔加入100μl包被液，4℃孵育过夜。包被液的制备方法：用pH9.0、0.05M碳酸盐包被缓冲液将重组人HER2蛋白/ErbB2蛋白(义翘神州，货号：10925-HCCH-50，命名为HER2-ECD)稀释为蛋白含量为2ng/μl的溶液。Take the ELISA plate, add 100 μl of coating solution to each well, and incubate overnight at 4°C. Preparation method of coating solution: Dilute recombinant human HER2 protein/ErbB2 protein (Shenzhou Yiqiao, product number: 10925-HCCH-50, named HER2-ECD) with pH 9.0, 0.05M carbonate coating buffer to A solution with a protein content of 2ng/μl.

2、封闭2. Closed

完成步骤1后，取所述酶标板，用PBST洗板3次(每次350μl，每次3分钟)，然后每孔加入200μl的10％小牛血清，37℃孵育2小时。After completing step 1, take the microplate and wash the plate 3 times with PBST (350 μl each time, 3 minutes each time), then add 200 μl of 10% calf serum to each well, and incubate at 37° C. for 2 hours.

3、加样3. Add sample

完成步骤2后，取所述酶标板，每孔加入100μl待测样品，37℃孵育2小时，然后PBST洗板3次(每次350μl，每次3分钟)。After completing step 2, take the ELISA plate, add 100 μl of the sample to be tested to each well, incubate at 37° C. for 2 hours, and then wash the plate 3 times with PBST (350 μl each time, 3 minutes each time).

待测样品为阳性对照抗体溶液(Trastuzumab溶液)或H5L5溶液。待测样本的蛋白浓度为0.0625μg/ml或0.03125μg/ml。The sample to be tested is positive control antibody solution (Trastuzumab solution) or H5L5 solution. The protein concentration of the sample to be tested is 0.0625 μg/ml or 0.03125 μg/ml.

4、加酶标抗体4. Add enzyme-labeled antibody

完成步骤3后，取所述酶标板，每孔加入100μl酶标抗体(山羊抗人IgG-HRP，中杉金桥，货号ZB-2304)，37℃孵育2小时，然后PBST洗板3次(每次350μl，每次3分钟)。After completing step 3, take the enzyme-labeled plate, add 100 μl enzyme-labeled antibody (goat anti-human IgG-HRP, Zhongshan Jinqiao, Cat. No. ZB-2304) to each well, incubate at 37°C for 2 hours, and then wash the plate 3 times with PBST (each 350 μl each time, 3 minutes each time).

5、显色5. Color rendering

完成步骤4后，取所述酶标板，每孔加入100μl TMB底物溶液(天根生物，PA107-01)，37℃孵育15分钟。After completing step 4, take the microtiter plate, add 100 μl of TMB substrate solution (Tiangen Biology, PA107-01) to each well, and incubate at 37° C. for 15 minutes.

6、终止反应6. Termination reaction

完成步骤5后，取所述酶标板，每孔加入50μl 2M硫酸溶液终止酶联反应。After step 5 is completed, the enzyme-linked reaction is terminated by adding 50 μl of 2M sulfuric acid solution to each well of the microtiter plate.

7、结果判定7. Result judgment

将酶标仪(MD190)扫描波长调至OD450，检测各孔OD值，若大于规定的阴性对照OD值的2.1倍，即为阳性。Adjust the scanning wavelength of the microplate reader (MD190) to OD450, and detect the OD value of each well. If it is greater than 2.1 times the OD value of the specified negative control, it is positive.

结果见图3。浓度为0.0625μg/ml和0.03125μg/ml的抗体H5L5与HER2-ECD在OD450均值分别为1.668与0.8169，浓度为0.0625μg/ml和0.03125μg/ml的对照抗体Trastuzumab与HER2-ECD在OD450值分别为1.657与1.123，结果表明H5L5和对照抗体Trastuzumab与HER2-ECD的亲和力相当。The results are shown in Figure 3. The average OD450 values of the antibodies H5L5 and HER2-ECD at concentrations of 0.0625 μg/ml and 0.03125 μg/ml were 1.668 and 0.8169, respectively, and the OD450 values of the control antibodies Trastuzumab and HER2-ECD at concentrations of 0.0625 μg/ml and 0.03125 μg/ml were respectively 1.657 and 1.123, the results show that H5L5 and the control antibody Trastuzumab have similar affinity to HER2-ECD.

二、流式细胞术检测2. Flow cytometry detection

1、取对数期生长的SK-BR-3细胞，制备成浓度为5×10⁵细胞/ml的细胞悬液。1. Take SK-BR-3 cells growing in logarithmic phase, and prepare a cell suspension with a concentration of 5×10 ⁵ cells/ml.

2、取1个EP管，加入1ml步骤1制备的细胞悬液，1000rpm低温(4℃)离心5min,弃上清，用1ml含1％FBS的PBS洗涤一次。2. Take an EP tube, add 1ml of the cell suspension prepared in step 1, centrifuge at 1000rpm at low temperature (4°C) for 5min, discard the supernatant, and wash once with 1ml of PBS containing 1% FBS.

3、完成步骤2后，取所述EP管，加入200μl待测溶液，4℃孵育60min，然后1000rpm低温(4℃)离心5min，弃上清，用500μl含1％FBS的PBS洗涤一次。3. After completing step 2, take the EP tube, add 200 μl of the solution to be tested, incubate at 4° C. for 60 minutes, then centrifuge at 1000 rpm at low temperature (4° C.) for 5 minutes, discard the supernatant, and wash once with 500 μl PBS containing 1% FBS.

待测溶液为Human IgG抗体溶液或阳性对照抗体溶液(Trastuzumab溶液)或H5L5溶液。待测溶液的蛋白浓度为5μg/ml。The solution to be tested is Human IgG antibody solution or positive control antibody solution (Trastuzumab solution) or H5L5 solution. The protein concentration of the solution to be tested was 5 μg/ml.

4、完成步骤3后，取所述EP管，加入100μlFITC标记的羊抗人IgG(GAH-FITC，1：32稀释)，4℃避光孵育45min。4. After completing step 3, take the EP tube, add 100 μl FITC-labeled goat anti-human IgG (GAH-FITC, diluted 1:32), and incubate at 4°C in the dark for 45 minutes.

5、完成步骤4后，取所述EP管，1000rpm低温(4℃)离心5min，弃上清，用500μl含1％FBS的PBS洗涤一次。5. After completing step 4, take the EP tube, centrifuge at 1000 rpm at low temperature (4° C.) for 5 min, discard the supernatant, and wash once with 500 μl of PBS containing 1% FBS.

6、完成步骤5后，取所述EP管，用300μl含1％FBS的PBS重悬细胞后流式检测。6. After completing step 5, take the EP tube, resuspend the cells in 300 μl PBS containing 1% FBS, and perform flow cytometric detection.

结果见图4。图4A为Human IgG的检测结果，图4B为对照抗体Trastuzumab的检测结果，图4C为抗体H5L5的检测结果。抗体H5L5与SK-BR-3细胞结合的阳性率为99.98％，平均荧光强度为27.04。对照抗体Trastuzumab与SK-BR-3结合的阳性率为99.60％，平均荧光强度为16.07。流式结果表明抗体H5L5可有效的结合SK-BR-3细胞，且与SK-BR-3细胞结合显著高于对照抗体Trastuzumab。The results are shown in Figure 4. Figure 4A is the detection result of Human IgG, Figure 4B is the detection result of the control antibody Trastuzumab, and Figure 4C is the detection result of the antibody H5L5. The positive rate of antibody H5L5 binding to SK-BR-3 cells was 99.98%, and the average fluorescence intensity was 27.04. The positive rate of the control antibody Trastuzumab binding to SK-BR-3 was 99.60%, and the average fluorescence intensity was 16.07. The results of flow cytometry showed that antibody H5L5 could effectively bind to SK-BR-3 cells, and the binding to SK-BR-3 cells was significantly higher than that of the control antibody Trastuzumab.

实施例4、细胞凋亡检测Embodiment 4, cell apoptosis detection

1、将SK-BR-3细胞按照5×10⁵细胞/孔的浓度接种6孔板，放入37℃、5％CO₂培养箱中培养24h。1. Inoculate SK-BR-3 cells in a 6-well plate at a concentration of 5×10 ⁵ cells/well, and culture them in a 37°C, 5% CO ₂ incubator for 24 hours.

2、完成步骤1后，取所述六孔板，将待测样品加至孔中，继续37℃、5％CO₂培养箱中培养36h。2. After step 1 is completed, take the six-well plate, add the sample to be tested into the wells, and continue culturing in a 37° C., 5% CO ₂ incubator for 36 hours.

待测样品为阳性对照抗体溶液或H5L5溶液，待测样品加至孔中后的蛋白浓度为10μg/ml。设置PBS替代待测样品的对照。The sample to be tested is positive control antibody solution or H5L5 solution, and the protein concentration after the sample to be tested is added to the well is 10 μg/ml. Set PBS to replace the control of the sample to be tested.

3、完成步骤2后，取所述六孔板，收集细胞后用1×Binding Buffer重悬细胞，加入5μl Annexin V-FITC室温孵育10min，然后加入5μlPI室温孵育10min，最后用300μl的1×Binding Buffer重悬细胞，进行流式上机检测分析。3. After completing step 2, take the six-well plate, collect the cells, resuspend the cells with 1×Binding Buffer, add 5 μl Annexin V-FITC and incubate at room temperature for 10 minutes, then add 5 μl PI and incubate at room temperature for 10 minutes, and finally use 300 μl of 1×Binding The cells were resuspended in Buffer and analyzed by flow cytometry.

结果如图5所示。图5A为PBS检测结果，图5B为Trastuzumab检测结果，图5C为H5L5检测结果。抗HER2抗体H5L5与阴性对照(PBS)相比差异显著，可使肿瘤细胞SK-BR-3发生凋亡，且诱导SK-BR-3细胞凋亡情况与Trastuzumab相当。The result is shown in Figure 5. Fig. 5A is the detection result of PBS, Fig. 5B is the detection result of Trastuzumab, and Fig. 5C is the detection result of H5L5. Compared with the negative control (PBS), the anti-HER2 antibody H5L5 has a significant difference, and can induce apoptosis of tumor cells SK-BR-3, and induces apoptosis of SK-BR-3 cells comparable to that of Trastuzumab.

实施例5、竞争性抑制实验Embodiment 5, competitive inhibition experiment

1、将Trastuzumab溶液(蛋白含量为1mg/ml)用透析反应液(7.56g NaHCO₃，1.06gNa₂CO₃，7.36g NaCl，加水定容至1L，pH＝9.0)进行透析，得到抗体溶液。1. Dialyze the Trastuzumab solution (protein content: 1 mg/ml) with the dialysis reaction solution (7.56g NaHCO ₃ , 1.06g Na ₂ CO ₃ , 7.36g NaCl, add water to 1L, pH=9.0) to obtain an antibody solution.

2、将FITC采用DMSO配制配置为浓度为1mg/ml的FITC溶液。2. FITC was prepared in DMSO to form a FITC solution with a concentration of 1 mg/ml.

3、按照蛋白质：FITC＝1mg:0.15mg的比例将FITC溶液缓慢加入于抗体溶液中，4℃避光反应8h，加入NH₄Cl至终浓度50mmol/L终止反应2h，透析袋中将多余FITC透析出来，最后进行交联物的测定，蛋白浓度(mg/ml)＝[A280–0.31×A495]/1.4；FITC/蛋白质：3.1×A495/[A280–0.31×A495]，FITC/蛋白质应介于2.5～6.5之间，得到FITC标记的Trastuzumab溶液。3. According to the ratio of protein:FITC=1mg:0.15mg, slowly add FITC solution to the antibody solution, react in the dark at 4°C for 8 hours, add NH ₄ Cl to a final concentration of 50mmol/L to stop the reaction for 2 hours, and put excess FITC in the dialysis bag After dialysis, the cross-linked product was finally determined, protein concentration (mg/ml) = [A280–0.31×A495]/1.4; FITC/protein: 3.1×A495/[A280–0.31×A495], FITC/protein should be mediated Between 2.5 and 6.5, a FITC-labeled Trastuzumab solution was obtained.

4、在EP管中，加入浓度为3μg/ml的FITC标记的Trastuzumab溶液和待测抗体溶液，冰上孵育1h。4. In the EP tube, add the FITC-labeled Trastuzumab solution and the antibody solution to be tested at a concentration of 3 μg/ml, and incubate on ice for 1 hour.

待测抗体溶液为Human IgG抗体溶液或阳性对照抗体溶液(Trastuzumab溶液)或H5L5溶液。待测抗体的浓度分别为200μg/ml、40μg/ml、8μg/ml、1.6μg/ml、0.32μg/ml、0.064μg/ml及0μg/ml。The antibody solution to be tested is Human IgG antibody solution or positive control antibody solution (Trastuzumab solution) or H5L5 solution. The concentrations of the antibodies to be tested were 200 μg/ml, 40 μg/ml, 8 μg/ml, 1.6 μg/ml, 0.32 μg/ml, 0.064 μg/ml and 0 μg/ml, respectively.

5、完成步骤4后，4℃离心5min弃上清，用含1％FBS的PBS洗涤一次。然后用300μl含1％FBS的PBS重悬细胞，流式检测。5. After completing step 4, centrifuge at 4°C for 5 minutes to discard the supernatant, and wash once with PBS containing 1% FBS. Then the cells were resuspended with 300 μl PBS containing 1% FBS, and flow cytometry was performed.

结果如图6所示。对照抗体Trastuzumab在1.6μg/ml就可以看到可以竞争性抑制Trastuzumab-FITC与SK-BR-3细胞的结合，而抗体H5L5在200μg/ml时也无法竞争性抑制Trastuzumab-FITC与SK-BR-3的结合，结合上述的亲和力情况，可以推测抗体H5L5与Trastuzumab具有不同的HER2抗原表位。The result is shown in Figure 6. The control antibody Trastuzumab can competitively inhibit the binding of Trastuzumab-FITC and SK-BR-3 cells at 1.6 μg/ml, while the antibody H5L5 cannot competitively inhibit the binding of Trastuzumab-FITC and SK-BR-3 cells at 200 μg/ml. 3, combined with the above-mentioned affinity, it can be speculated that the antibody H5L5 and Trastuzumab have different HER2 epitopes.

序列表 sequence listing

<110> 中国人民解放军军事科学院军事医学研究院<110> Academy of Military Medical Sciences, Chinese People's Liberation Army

安徽大学 Anhui University

<120> 抗HER2抗原的人源化抗体H5L5及其应用<120> Humanized antibody H5L5 against HER2 antigen and its application

<160> 8<160> 8

<170> SIPOSequenceListing 1.0<170> SIP Sequence Listing 1.0

<210> 1<210> 1

<211> 469<211> 469

<212> PRT<212> PRT

<213> 人工序列(Artificial Sequence)<213> Artificial Sequence

<400> 1<400> 1

Met Asp Trp Thr Trp Arg Val Phe Cys Leu Leu Ala Val Ala Pro GlyMet Asp Trp Thr Trp Arg Val Phe Cys Leu Leu Ala Val Ala Pro Gly

1 5 10 151 5 10 15

Ala Gln Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val GlnAla Gln Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln

20 25 30 20 25 30

Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn IlePro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile

35 40 45 35 40 45

Lys Asp Thr Tyr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly LeuLys Asp Thr Tyr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

50 55 60 50 55 60

Glu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr AlaGlu Trp Val Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala

65 70 75 8065 70 75 80

Asp Ser Val Lys Gly Arg Phe Ser Ile Ser Ala Asp Asn Ser Lys AsnAsp Ser Val Lys Gly Arg Phe Ser Ile Ser Ala Asp Asn Ser Lys Asn

85 90 95 85 90 95

Thr Leu Tyr Leu Glu Met Asn Ser Leu Arg Ala Glu Asp Thr Ala ValThr Leu Tyr Leu Glu Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val

100 105 110 100 105 110

Tyr Tyr Cys Ala Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp TyrTyr Tyr Cys Ala Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr

115 120 125 115 120 125

Trp Gly Gln Gly Ala Leu Val Thr Val Ser Ser Ala Ser Thr Lys GlyTrp Gly Gln Gly Ala Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly

130 135 140 130 135 140

Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly GlyPro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly

145 150 155 160145 150 155 160

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro ValThr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

165 170 175 165 170 175

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr PheThr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

180 185 190 180 185 190

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val ValPro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Ser Val Val

195 200 205 195 200 205

Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn ValThr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val

210 215 220 210 215 220

Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro LysAsn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys

225 230 235 240225 230 235 240

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu LeuSer Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

245 250 255 245 250 255

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp ThrLeu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

260 265 270 260 265 270

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp ValLeu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

275 280 285 275 280 285

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly ValSer His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

290 295 300 290 295 300

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn SerGlu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

305 310 315 320305 310 315 320

Thr Tyr Arg Val Val Ser Val Leu Ser Val Leu His Gln Asp Trp LeuThr Tyr Arg Val Val Ser Val Leu Ser Val Leu His Gln Asp Trp Leu

325 330 335 325 330 335

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro AlaAsn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

340 345 350 340 345 350

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu ProPro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

355 360 365 355 360 365

Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn GlnGln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln

370 375 380 370 375 380

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile AlaVal Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

385 390 395 400385 390 395 400

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr ThrVal Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

405 410 415 405 410 415

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys LeuPro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

420 425 430 420 425 430

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys SerThr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

435 440 445 435 440 445

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu SerVal Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

450 455 460 450 455 460

Leu Ser Pro Gly LysLeu Ser Pro Gly Lys

465465

<210> 2<210> 2

<211> 1429<211> 1429

<212> DNA<212>DNA

<213> 人工序列(Artificial Sequence)<213> Artificial Sequence

<400> 2<400> 2

aagcttaatt gccgccacca tggactggac ctggagggtc ttctgcttgc tggctgtagc 60aagcttaatt gccgccacca tggactggac ctggagggtc ttctgcttgc tggctgtagc 60

tccaggtgct cactcccagg ttcagctggt ggagtctggc ggtggcgtgg tgcagccagg 120tccaggtgct cactcccagg ttcagctggt ggagtctggc ggtggcgtgg tgcagccagg 120

gcggtcactc cgtttgtcct gtgcagcttc tggcttcaac attaaagaca cctatatgca 180gcggtcactc cgtttgtcct gtgcagcttc tggcttcaac attaaagaca cctatatgca 180

ctgggtgcgt caggccccgg gtaagggcct ggaatgggtt gcaaggattt atcctacgaa 240ctgggtgcgt caggccccgg gtaagggcct ggaatgggtt gcaaggattt atcctacgaa 240

tggttatact agatatgccg atagcgtcaa gggccgtttc agcataagcg cagacaactc 300tggttatact agatatgccg atagcgtcaa gggccgtttc agcataagcg cagacaactc 300

caaaaacaca ctgtacctgg agatgaacag cctgcgtgct gaggacactg ccgtctatta 360caaaaacaca ctgtacctgg agatgaacag cctgcgtgct gaggacactg ccgtctatta 360

ttgtgccaga tggggagggg acggcttcta tgctatggac tactggggtc aaggagccct 420ttgtgccaga tggggagggg acggcttcta tgctatggac tactggggtc aaggagccct 420

ggtcaccgtc tcctcggcta gcaccaaggg cccatcggtc ttccccctgg caccctcctc 480ggtcaccgtc tcctcggcta gcaccaaggg cccatcggtc ttccccctgg caccctcctc 480

caagagcacc tctgggggca cagcggccct gggctgcctg gtcaaggact acttccccga 540caagagcacc tctgggggca cagcggccct gggctgcctg gtcaaggact acttccccga 540

accggtgacg gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc 600accggtgacg gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc 600

tgtcctacag tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag 660tgtcctacag tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag 660

cttgggcacc cagacctaca tctgcaacgt gaatcacaag cccagcaaca ccaaggtgga 720cttgggcacc cagacctaca tctgcaacgt gaatcacaag cccagcaaca ccaaggtgga 720

caagaaagtt gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc 780caagaaagtt gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc 780

tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat 840tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat 840

gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga 900gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga 900

ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg 960ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg 960

ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctctccgtcc tgcaccagga 1020ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctctccgtcc tgcaccagga 1020

ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat 1080ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat 1080

cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc 1140cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc 1140

tccatctcgg gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt 1200tccatctcgg gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt 1200

ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa 1260ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa 1260

gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctatagca agctcaccgt 1320gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctatagca agctcaccgt 1320

ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct 1380ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct 1380

gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatga 1429gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatga 1429

<210> 3<210> 3

<211> 234<211> 234

<212> PRT<212> PRT

<213> 人工序列(Artificial Sequence)<213> Artificial Sequence

<400> 3<400> 3

Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu ProMet Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Leu Trp Leu Pro

1 5 10 151 5 10 15

Asp Thr Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu SerAsp Thr Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Ser Leu Ser

20 25 30 20 25 30

Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln AspAla Ser Val Gly Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp

35 40 45 35 40 45

Ile Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProIle Asn Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

50 55 60 50 55 60

Lys Leu Leu Ile Tyr Ser Ala Ser Phe Asp Tyr Ser Gly Val Pro SerLys Leu Leu Ile Tyr Ser Ala Ser Phe Asp Tyr Ser Gly Val Pro Ser

65 70 75 8065 70 75 80

Arg Phe Ser Gly Ser Gly Ser Gly Arg Asp Phe Thr Phe Thr Ile SerArg Phe Ser Gly Ser Gly Ser Gly Arg Asp Phe Thr Phe Thr Ile Ser

85 90 95 85 90 95

Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln His TyrSer Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys Gln Gln His Tyr

100 105 110 100 105 110

Thr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys ArgThr Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg

115 120 125 115 120 125

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu GlnThr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

130 135 140 130 135 140

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe TyrLeu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Asn Phe Tyr

145 150 155 160145 150 155 160

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln SerPro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

165 170 175 165 170 175

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser ThrGly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

180 185 190 180 185 190

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu LysTyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

195 200 205 195 200 205

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser ProHis Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

210 215 220 210 215 220

Val Thr Lys Ser Phe Asn Arg Gly Glu CysVal Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230225 230

<210> 4<210> 4

<211> 724<211> 724

<212> DNA<212>DNA

<213> 人工序列(Artificial Sequence)<213> Artificial Sequence

<400> 4<400> 4

aagcttaatt gccgccacca tggagacacc cgcccagctg ctgttcctgc tgctgctgtg 60aagcttaatt gccgccacca tggagacacc cgcccagctg ctgttcctgc tgctgctgtg 60

gctgcccgac accaccggcg acatccagat gacccagtcc ccgagctccc tgtccgcctc 120gctgcccgac accaccggcg acatccagat gacccagtcc ccgagctccc tgtccgcctc 120

tgtgggcgat agggttacca tcacctgcca ggccagtcag gatatcaata ctgctgtagc 180tgtgggcgat agggttacca tcacctgcca ggccagtcag gatatcaata ctgctgtagc 180

ctggtatcaa cagaaaccag gaaaagctcc gaaactactg atttactcgg catccttcga 240ctggtatcaa cagaaaccag gaaaagctcc gaaactactg atttactcgg catccttcga 240

ctactctgga gtcccttctc gcttctctgg ctccggatct gggagggatt tcactttcac 300ctactctgga gtcccttctc gcttctctgg ctccggatct gggagggatt tcactttcac 300

catcagcagt ctgcagccgg aagacgtggc aacttattac tgtcagcaac attatactac 360catcagcagt ctgcagccgg aagacgtggc aacttattac tgtcagcaac attatactac 360

tcctcccacg ttcggacagg gtaccaaggt ggagatcaag cggaccgtgg cggcgccatc 420tcctccacg ttcggacagg gtaccaaggt ggagatcaag cggaccgtgg cggcgccatc 420

tgtcttcatc ttcccgccat ctgatgagca gttgaaatct ggtaccgcta gcgttgtgtg 480tgtcttcatc ttcccgccat ctgatgagca gttgaaatct ggtaccgcta gcgttgtgtg 480

cctgctgaat aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct 540cctgctgaat aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct 540

ccaatcgggt aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag 600ccaatcgggt aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag 600

cctcagcagc accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg 660cctcagcagc accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg 660

cgaagtcacc catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg 720cgaagtcacc catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg 720

ttag 724ttag 724

<210> 5<210> 5

<211> 120<211> 120

<212> PRT<212> PRT

<213> 人工序列(Artificial Sequence)<213> Artificial Sequence

<400> 5<400> 5

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly GlyGlu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 151 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp ThrSer Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr

20 25 30 20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp ValTyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45 35 40 45

Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser ValAla Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val

50 55 60 50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala TyrLys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 8065 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr CysLeu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95 85 90 95

Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly GlnSer Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110 100 105 110

Gly Thr Leu Val Thr Val Ser SerGly Thr Leu Val Thr Val Ser Ser

115 120 115 120

<210> 6<210> 6

<211> 108<211> 108

<212> PRT<212> PRT

<213> 人工序列(Artificial Sequence)<213> Artificial Sequence

<400> 6<400> 6

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val GlyAsp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 151 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr AlaAsp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala

20 25 30 20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu IleVal Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45 35 40 45

Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser GlyTyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60 50 55 60

Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln ProSer Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 8065 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro ProGlu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro

85 90 95 85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys ArgThr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg

100 105 100 105

<210> 7<210> 7

<211> 360<211> 360

<212> DNA<212>DNA

<213> 人工序列(Artificial Sequence)<213> Artificial Sequence

<400> 7<400> 7

gaggttcagc tggtggagtc tggcggtggc ctggtgcagc cagggggctc actccgtttg 60gaggttcagc tggtggagtc tggcggtggc ctggtgcagc cagggggctc actccgtttg 60

tcctgtgcag cttctggctt caacattaaa gacacctata tacactgggt gcgtcaggcc 120tcctgtgcag cttctggctt caacattaaa gacacctata tacactgggt gcgtcaggcc 120

ccgggtaagg gcctggaatg ggttgcaagg atttatccta cgaatggtta tactagatat 180ccgggtaagg gcctggaatg ggttgcaagg atttatccta cgaatggtta tactagatat 180

gccgatagcg tcaagggccg tttcactata agcgcagaca catccaaaaa cacagcctac 240gccgatagcg tcaagggccg tttcactata agcgcagaca catccaaaaa cacagcctac 240

ctgcagatga acagcctgcg tgctgaggac actgccgtct attattgttc tagatgggga 300ctgcagatga acagcctgcg tgctgaggac actgccgtct attattgttc tagatgggga 300

ggggacggct tctatgctat ggactactgg ggtcaaggaa ccctggtcac cgtctcctcg 360ggggacggct tctatgctat ggactactgg ggtcaaggaa ccctggtcac cgtctcctcg 360

<210> 8<210> 8

<211> 324<211> 324

<212> DNA<212>DNA

<213> 人工序列(Artificial Sequence)<213> Artificial Sequence

<400> 8<400> 8

gacatccaga tgacccagtc cccgagctcc ctgtccgcct ctgtgggcga tagggttacc 60gacatccaga tgacccagtc cccgagctcc ctgtccgcct ctgtgggcga tagggttacc 60

atcacctgcc gtgccagtca ggatgtgaat actgctgtag cctggtatca acagaaacca 120atcacctgcc gtgccagtca ggatgtgaat actgctgtag cctggtatca acagaaacca 120

ggaaaagctc cgaaactact gatttactcg gcatccttcc tctactctgg agtcccttct 180ggaaaagctc cgaaactact gatttactcg gcatccttcc tctactctgg agtcccttct 180

cgcttctctg gctccagatc tgggacggat ttcactctga ccatcagcag tctgcagccg 240cgcttctctg gctccagatc tgggacggat ttcactctga ccatcagcag tctgcagccg 240

gaagacttcg caacttatta ctgtcagcaa cattatacta ctcctcccac gttcggacag 300gaagacttcg caacttatta ctgtcagcaa catttatacta ctcctcccac gttcggacag 300

ggtaccaagg tggagatcaa gcgg 324ggtaccaagg tggagatcaa gcgg 324

Claims (10)

1. a kind of IgG, CDR1, CDR2 and CDR3 in heavy chain variable region are successively if the sequence 1 of sequence table is from N-terminal 50-54 Shown in amino acids residue, 69-85 amino acids residue and 118-128 amino acids residues, in light chain variable region CDR1, CDR2 and CDR3 are successively as the sequence 3 of sequence table is residual from N-terminal 44-54 amino acids residue, 70-76 amino acids Shown in base and 109-117 amino acids residues.

2. IgG as described in claim 1, it is characterised in that：

The sequence 1 of the heavy chain variable region such as sequence table is from shown in N-terminal 20-139 amino acids residues；

The sequence 3 of the light chain variable region such as sequence table is from shown in N-terminal 21-128 amino acids residues.

3. IgG as claimed in claim 2, it is characterised in that：

The heavy chain is following (a1) or (a2)：(a1) sequence 1 of sequence table is formed from N-terminal 20-469 amino acids residues Protein；(a2) protein shown in the sequence 1 of sequence table；

The light chain is following (b1) or (b2)：(b1) sequence 3 of sequence table is formed from N-terminal 21-234 amino acids residues Protein；(b2) protein shown in the sequence 3 of sequence table.

4. encoding the gene of any IgG of claims 1 to 3.

5. gene as claimed in claim 4, it is characterised in that：

The gene for encoding the heavy chain is following (c1) or (c2) or (c3)：

(c1) sequence 2 of sequence table holds DNA molecular shown in 77-1426 nucleotide from 5 '；

(c2) sequence 2 of sequence table holds DNA molecular shown in 20-1429 nucleotide from 5 '；

(c3) DNA molecular shown in the sequence 2 of sequence table；

The gene for encoding the light chain is following (d1) or (d2) or (d3)：

(d1) sequence 4 of sequence table holds DNA molecular shown in 80-821 nucleotide from 5 '；

(d2) sequence 4 of sequence table holds DNA molecular shown in 20-724 nucleotide from 5 '；

(d3) DNA molecular shown in the sequence 4 of sequence table.

6. applications of any IgG of claims 1 to 3 in preparing the drug for inducing apoptosis of tumour cell.

7. applications of any IgG of claims 1 to 3 in preparing the drug for treating tumour.

8. applications of any IgG of claims 1 to 3 in preparing product；The purposes of the product be following (f1) or (f2) or (f3) or (f4)：

(f1) tumour cell is detected；

(f2) tumour cell is combined；

(f3) human epidermal growth factor receptor 2 is detected；

(f4) human epidermal growth factor receptor 2 is combined.

9. a kind of product, active constituent is any IgG of claims 1 to 3；The purposes of the product be following (f1) or (f2) or (f3) or (f4)：

(f1) tumour cell is detected；

(f2) tumour cell is combined；

(f3) human epidermal growth factor receptor 2 is detected；

(f4) human epidermal growth factor receptor 2 is combined.

10. the application as described in claim 6 to 8 is any, or, the product described in claim 9, it is characterised in that：It is described swollen Oncocyte is breast cancer cell；The tumour is breast cancer.

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Cited By (2)

* Cited by examiner, † Cited by third party

Publication number	Priority date	Publication date	Assignee	Title
CN109912716A (en) *	2017-12-13	2019-06-21	凯惠科技发展(上海)有限公司	A kind of EGFR antibody and its preparation method and application
CN112391414A (en) *	2020-11-17	2021-02-23	中国医学科学院基础医学研究所	HER 2-targeted CAR-T expression vector and construction and application thereof

Citations (5)

* Cited by examiner, † Cited by third party

Publication number	Priority date	Publication date	Assignee	Title
CN103153339A (en) *	2010-05-27	2013-06-12	根马布股份公司	Monoclonal antibodies against her2 epitope
CN103936857A (en) *	2014-03-27	2014-07-23	中国人民解放军军事医学科学院生物工程研究所	Antibody L5H5 with CD20-resistant antigen and application thereof
CN104497140A (en) *	2014-11-26	2015-04-08	嘉和生物药业有限公司	Fully humanized HER2 antibody as well as encoding gene and application of antibody
CN104650228A (en) *	2014-11-26	2015-05-27	嘉和生物药业有限公司	Full humanized HER2 antibody as well as coding gene and application thereof
CN107074962A (en) *	2014-06-18	2017-08-18	梅尔莎纳医疗公司	For the monoclonal antibody and its application method of HER2 epitopes

2018
- 2018-04-11 CN CN201810320106.8A patent/CN108484772A/en active Pending

Patent Citations (5)

* Cited by examiner, † Cited by third party

Publication number	Priority date	Publication date	Assignee	Title
CN103153339A (en) *	2010-05-27	2013-06-12	根马布股份公司	Monoclonal antibodies against her2 epitope
CN103936857A (en) *	2014-03-27	2014-07-23	中国人民解放军军事医学科学院生物工程研究所	Antibody L5H5 with CD20-resistant antigen and application thereof
CN107074962A (en) *	2014-06-18	2017-08-18	梅尔莎纳医疗公司	For the monoclonal antibody and its application method of HER2 epitopes
CN104497140A (en) *	2014-11-26	2015-04-08	嘉和生物药业有限公司	Fully humanized HER2 antibody as well as encoding gene and application of antibody
CN104650228A (en) *	2014-11-26	2015-05-27	嘉和生物药业有限公司	Full humanized HER2 antibody as well as coding gene and application thereof

Cited By (3)

* Cited by examiner, † Cited by third party

Publication number	Priority date	Publication date	Assignee	Title
CN109912716A (en) *	2017-12-13	2019-06-21	凯惠科技发展(上海)有限公司	A kind of EGFR antibody and its preparation method and application
CN109912716B (en) *	2017-12-13	2022-08-23	凯惠科技发展(上海)有限公司	EGFR antibody and preparation method and application thereof
CN112391414A (en) *	2020-11-17	2021-02-23	中国医学科学院基础医学研究所	HER 2-targeted CAR-T expression vector and construction and application thereof

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CN108484772A - The humanized antibody H5L5 of anti-HER2 antigens and its application - Google Patents