WO2021056907A1 - 杂交瘤细胞lcz9h4及其分泌的单克隆抗体和应用 - Google Patents

杂交瘤细胞lcz9h4及其分泌的单克隆抗体和应用 Download PDF

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WO2021056907A1
WO2021056907A1 PCT/CN2019/129882 CN2019129882W WO2021056907A1 WO 2021056907 A1 WO2021056907 A1 WO 2021056907A1 CN 2019129882 W CN2019129882 W CN 2019129882W WO 2021056907 A1 WO2021056907 A1 WO 2021056907A1
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kit
lcz9h4
monoclonal antibody
hybridoma cell
catenin
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钱泽
孟洁
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昂科生物医学技术(苏州)有限公司
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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
    • C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
    • 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
    • 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
    • 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
    • G01N33/57407—Specifically defined cancers
    • G01N33/57434—Specifically defined cancers of prostate
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    • G01—MEASURING; TESTING
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    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • 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
    • 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
    • G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/82—Translation products from oncogenes

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  • the invention belongs to the technical field of clinical diagnosis, and particularly relates to a hybridoma cell LCZ9H4 and a preparation method thereof, and the application of the monoclonal antibody and the monoclonal antibody secreted by the hybridoma cell LCZ9H4 in a kit for detecting human prostate cancer.
  • prostate cancer The treatment and prognosis of prostate cancer mainly depend on early diagnosis. Although there is no mandatory requirement to test men who are generally at risk of prostate cancer, the American Cancer Society recommends that men start prostate-specific antigen (PSA) testing and digital rectal examination (DRE) from the age of 50. Men who are at a higher risk of prostate cancer should start testing at the age of 45. In the past few decades, due to the early screening of prostate cancer, prostate cancer was detected early in the initial and local development stages. The 5-year survival rate of prostate cancer patients has increased significantly, which also fully demonstrates the early detection of prostate cancer. The importance of early diagnosis.
  • PSA prostate-specific antigen
  • DRE digital rectal examination
  • prostate specific antigen PSA
  • fPSA/tPSA DRE
  • DRE digital rectal examination
  • prostate cancer biomarkers to supplement PSA screening, which can improve the accuracy of prostate cancer risk assessment and predict its development trend, and avoid over-diagnosis and treatment of patients.
  • the development of prostate cancer biomarkers independent of PSA detection can not only improve the accuracy of early diagnosis, but also provide a good basis for treatment.
  • the main technical problem solved by the present invention is to provide a hybridoma cell LCZ9H4 and a preparation method thereof, a monoclonal antibody secreted by the hybridoma cell and a kit for detecting human prostate cancer.
  • the present invention adopts the following technical solutions:
  • a hybridoma cell LCZ9H4 characterized in that: the hybridoma cell LCZ9H4 is deposited in the Chinese Type Culture Collection, and the deposit number is CCTCC No: C2019145.
  • the present invention also provides a monoclonal antibody, which is secreted and produced by the hybridoma cell LCZ9H4.
  • the present invention also provides a kit, which includes the hybridoma cell or the monoclonal antibody.
  • the kit is a chemiluminescence kit.
  • the invention also provides the use of the hybridoma cell or the monoclonal antibody in the preparation of a kit.
  • the kit is a kit based on immunoassay.
  • the kit is a chemiluminescence kit.
  • the kit is used for detecting human prostate cancer.
  • the present invention also provides the use of the hybridoma cell or the monoclonal antibody in preparing a kit for the diagnosis of human prostate cancer.
  • the hybridoma cells can stably produce anti-delta-catenin monoclonal antibodies, with stable chromosomes and high antibody titer, which can be used in the research of delta-catenin applications;
  • the anti- ⁇ -catenin monoclonal antibody can be used in the preparation of human prostate cancer diagnostic kits.
  • the test samples include human urine, blood, semen, prostate massage fluid and other human samples, which are used for the diagnosis of human prostate cancer. , which greatly makes up for the shortcomings of the existing clinical diagnosis technology.
  • the detection method of the present invention is not limited to chemiluminescence immunotechnology, but also includes colloidal gold immunochromatography, fluorescence immunochromatography and other similar immunoassay techniques.
  • Example 1 The preparation of hybridoma cell LCZ9H4 includes the following steps:
  • mouse anti-delta-catenin hybridoma cells includes the following steps:
  • Boost immunity For the first immunization, use 60ug/ml ⁇ -catenin protein plus Freund's complete adjuvant to emulsify the mixture in equal volume and then intramuscularly. Two weeks later, use 30ug/ml delta-catenin protein plus Freund’s incomplete adjuvant to boost the immunization twice with an interval of 2 weeks.
  • the serum antibody titer was detected by ELISA, and the titer of immunized mice was significantly above 1:100,000.
  • Prepare Boost immunity For the first immunization, use 60ug/ml ⁇ -catenin protein plus Freund's complete adjuvant to emulsify the mixture in equal volume and then intramuscularly. Two weeks later, use 30ug/ml delta-catenin protein plus Freund’s incomplete adjuvant to boost the immunization twice with an interval of 2 weeks.
  • the serum antibody titer was detected by ELISA, and the titer of immunized mice was significantly above 1:100,000.
  • the ELISA detection method is specifically: adding a coating buffer to a 96-well plate to dilute the ⁇ -catenin protein antigen to 3ug/ml, adding 50u1 to each reaction well for 1 hour, and washing 3 times with a washing solution. Add 100u1/well blocking solution to seal at 37°C for 2h, wash 3 times, and pat dry. Add the serum to be tested, dilute 1:800 in the first well, and dilute in a 1:2 gradient, incubate at 37°C for 30 minutes, wash the plate 4 times, and pat dry.
  • spleen cells and myeloma cells at a cell number of about 10:1, and fuse them under the fusion of polyethylene glycol (PEG, molecular weight 1500).
  • the selection medium is RPMI1640 medium containing HAT (50ml).
  • the positive hybridoma cells that can react with the delta-catenin protein were screened by indirect ELISA method, and the positive hybridoma cells obtained from the preliminary screening were expanded and cultured, and the labeled protein hybridoma cells were excluded to re-screen the target cells.
  • ⁇ -catenin protein instead of labeled hybridoma cells;
  • the indirect ELISA method was used to detect whether all five positive cells could specifically recognize ⁇ -catenin: a positive cell named A can specifically bind to ⁇ -catenin, indicating that A can specifically recognize ⁇ -catenin.
  • a hybridoma cell A that can stably secrete a monoclonal antibody against human delta-catenin was obtained.
  • the hybridoma cell A was classified and named as hybridoma cell LCZ9H4, which was deposited in China on August 28, 2019. The address is Wuhan University, China, and the deposit number is CCTCC N0:C2019145.
  • Example 2 Preparation and purification of mouse anti- ⁇ -catenin ascites monoclonal antibody, including the following steps:
  • mice choose healthy female BALB/C mice aged 8-12 weeks, weighing about 20g, and inject sterilized liquid paraffin, 0.5ml per mouse; 7 days later, inject 1 ⁇ 10 6 mice into each mouse.
  • the 5 monoclonal hybridoma cells of Example 1, LCZ9H4-1, LCZ9H4-2, LCZ9H4-3, LCZ9H4-4, LCZ9H4-5, each of the monoclonal hybridoma cells were injected with two mice;
  • UV spectroscopy is used to determine the absorbance values A280 and A260 of monoclonal antibodies at 280nm and 260nm. The result is a protein content of 21 mg/ml.
  • the kit uses chemiluminescence immunoassay.
  • the labeling buffer is a CB system, the pH is 19.5, the feed ratio (molar ratio) is 20:1 (acridine/antibody), and lysine is added when the reaction is terminated (the ratio of lysine/acridine is 140:1).
  • the labeled antibody is purified by Sephadex G-50 Sephadex column;

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Abstract

一种杂交瘤细胞LCZ9H4及其分泌的单克隆抗体和应用，该杂交瘤细胞LCZ9H4保藏于中国典型培养物保藏中心，保藏编号为CCTCC No：C2019145。该细胞能够稳定产生抗δ-catenin单克隆抗体，应用于δ-catenin的应用的研究中。

Description

杂交瘤细胞LCZ9H4及其分泌的单克隆抗体和应用 技术领域
本发明属于临床诊断技术领域，特别是涉及一种杂交瘤细胞LCZ9H4及其制备方法，杂交瘤细胞LCZ9H4分泌的单克隆抗体和单克隆抗体在检测人前列腺癌的试剂盒中的应用。
背景技术
前列腺癌的治疗及预后主要取决于早期诊断。虽然没有强制要求对一般有患前列腺癌风险的男性进行检测，但是美国癌症协会建议男性从50岁开始进行前列腺特异抗原(PSA)检测和直肠指检(DRE)。处于患前列腺癌风险较高的男性应在45岁开始检测。在过去的几十年间，由于前列腺癌的早期筛查使得前列腺癌在初期和局部发展阶段就被及早的检测出来，前列腺癌患者的5年存活率显著提高，这也充分表明了前列腺癌早发现早诊断的重要性。
临床上主要采用前列腺特异性抗原(PSA)、fPSA/tPSA和直肠指检(DRE)来筛查前列腺癌，最初可疑前列腺癌患者通常由直肠指检或血清PSA检查后再决定是否进行前列腺穿刺活检。前列腺癌患者主要通过前列腺系统性穿刺活检取得组织再病理学诊断得以确诊。
但是，大量的数据表明血清PSA水平的升高有可能是前列腺癌以外的因素导致，如良性前列腺增生。因此，假阳性是PSA筛查的主要弊端，并可能导致不必要的活检或其他令患者痛苦的干预治疗。另外，术前血清PSA水平并不总是与癌症体积或前列腺癌Gleason分数相关。此外相当一部分患者处于诊断灰区(血清tPSA介于4-10ng/ml，国内推荐fPSA/tPSA>0.16为正常参考值)，但是即使fPSA/tPSA>0.25，发生前列腺癌的可能性仍约为8％，使临床上相当部分患者不能被早期诊断。许多研究估计PSA检查无法检测出超过20％至30％的前列腺癌病患。最近，美国一些针对临床数据的调查得出的结论是PSA筛查无法很好地检测出前列腺癌。
因此，开发更可靠的生物标记物来补充PSA筛查就变得尤为重要，这样可以提高前列腺癌的风险评估和预测其发展趋势的准确性,避免对患者的过度诊 断和治疗。开发独立于PSA检测的前列腺癌生物标记物不仅可以提高早期诊断的准确性还可以为治疗方案提供很好的指导依据。
发明内容
本发明主要解决的技术问题是提供一种杂交瘤细胞LCZ9H4及其制备方法、该杂交瘤细胞分泌的单克隆抗体和检测人前列腺癌的试剂盒。
为解决上述技术问题，本发明采取了以下技术方案：
一种杂交瘤细胞LCZ9H4，其特征在于：所述杂交瘤细胞LCZ9H4保藏于中国典型培养物保藏中心，保藏编号为CCTCC No：C2019145。
本发明还提供一种单克隆抗体，所述单克隆抗体由所述的杂交瘤细胞LCZ9H4分泌产生。
本发明还提供一种试剂盒，所述试剂盒包括所述的杂交瘤细胞或所述的单克隆抗体。
进一步地说，所述试剂盒为化学发光试剂盒。
本发明还提供了所述的杂交瘤细胞或所述的单克隆抗体在制备试剂盒中的用途。
进一步地说，所述试剂盒是基于免疫检测的试剂盒。
进一步地说，所述试剂盒为化学发光试剂盒。
进一步地说，所述试剂盒用于检测人前列腺癌。
本发明还提供了所述的杂交瘤细胞或所述的单克隆抗体在制备用于人前列腺癌诊断的试剂盒中的用途。
本发明的有益效果至少具有以下几点：
1、该杂交瘤细胞能够稳定产生抗δ-catenin单克隆抗体，且细胞染色体稳定、抗体效价高，完全能够应用于δ-catenin的应用的研究中；
2、该抗δ-catenin的单克隆抗体可应用于人前列腺癌诊断试剂盒的制备中，检测样本包括了人体尿液、血液、精液、前列腺按摩液等人体样本，用于人前列腺癌的诊断，大大弥补现有临床诊断技术的不足。
3、本发明的检测方法不仅限于化学发光免疫技术，也包括了胶体金免疫层 析技术、荧光免疫层析技术等各种类似的免疫技术。
具体实施方式
下面对本发明的较佳实施例进行详细阐述，以使本发明的优点和特征能更易于被本领域技术人员理解，从而对本发明的保护范围做出更为清楚明确的界定。
实施例1：杂交瘤细胞LCZ9H4的制备，包括以下步骤：
1、鼠抗δ-catenin杂交瘤细胞的建立，包括以下步骤：
a.选用8只8-12周龄、体重20g左右且健康的雌性BALB/C小鼠，适用性饲养1周后，采集阴性血作为对照用；
b.初次免疫用60ug/ml δ-catenin蛋白加弗氏完全佐剂等体积搅拌乳化，肌肉注射。两周后，用30ug/ml δ-catenin蛋白加弗氏不完全佐剂强化免疫2次，每次间隔2周，ELISA检测血清抗体效价，免疫小鼠效价明显达到1:100000以上，准备加强免疫；
其中，ELISA检测法具体为：在96孔板中加入用包被缓冲液将δ-catenin蛋白抗原稀释至3ug/ml，在每个反应孔中加50u1，1小时，用洗液洗涤3次，加100u1/孔封闭液在37℃封闭2h，洗涤3次，拍干。加入待测血清，第一孔1:800稀释，往下1:2的梯度倍比稀释，37℃孵育30min，洗板4次，拍干。取辣根酶标记的羊抗鼠IgG按照1:2500倍稀释，100u1/孔加入反应孔，37℃孵育30min，洗涤4次，拍干。最后加入TMB 100u1/孔，显色15-30min。然后加入终止液，50u1/孔。以450nm单波长测定各孔OD值，并用阳性对照孔作为判断阳性。结果为血清效价为1:100000的阳性小鼠，可以用于细胞融合。
c.用40ug/ml δ-catenin蛋白肌肉注射强化免疫(不含佐剂)，3天后尾部采血，ELISA检测血清抗体效价，同时取脾脏。
d.将脾脏细胞与骨髓瘤细胞按细胞数10:1左右进行混合，并在聚乙二醇(PEG，分子量为1500)的促融作用下进行融合，选择培养基为含HAT的RPMI1640培养基(50ml)。11天后，通过间接ELISA方法筛选出能与δ-catenin蛋白反应的阳性杂交瘤细胞，并将初筛得到的阳性杂交瘤细胞扩大培养，并进行标 签蛋白杂交瘤细胞的排除，以复筛出针对δ-catenin蛋白而非标签的杂交瘤细胞；
e.用有限稀释法将获得的阳性杂交瘤细胞进行克隆，经过6次筛选最终确定5株能稳定分泌单克隆抗体的杂交瘤细胞A-1，A-2，A-3，A-4，A-5。
间接ELISA方法检测所有阳性5株细胞是否能够特异识别δ-catenin：一株命名为A的阳性细胞可以与δ-catenin特异结合，说明A可以特异识别δ-catenin。经筛选得到能稳定分泌抗人δ-catenin的单克隆抗体的杂交瘤细胞A，该杂交瘤细胞A的分类命名为杂交瘤细胞LCZ9H4，已于2019年8月28日保藏于保藏于中国典型培养物保藏中心，地址为中国武汉大学，保藏号为CCTCC N0:C2019145。
2、鼠抗人δ-catenin单克隆抗体的制备及单抗亚型的鉴定
取1000u1杂交瘤细胞LCZ9H4(5株杂交瘤细胞LCZ9H4-1，LCZ9H4-2，LCZ9H4-3，LCZ9H4-4，LCZ9H4-5)的培养上清，采用sigma分型试剂盒鉴定单克隆抗体的亚型，结果显示：单克隆抗体的重链为IgGl型，轻链为Kappa型。
实施例2鼠抗δ-catenin的腹水单克隆抗体制备及纯化，包括以下步骤：
a.选择8-12周龄、体重20g左右且健康的雌性BALB/C小鼠，腹腔注射灭菌液体石蜡，0.5m1/只；7天后，给每只小鼠腹腔注射1×10 6个实施例1的5株单克隆杂交瘤细胞LCZ9H4-1，LCZ9H4-2，LCZ9H4-3，LCZ9H4-4，LCZ9H4-5，每株单克隆杂交瘤细胞注射两只小鼠；
b.7天后观察小鼠腹水生产情况，如腹部明显膨大，即可抽取腹水。腹水采集后13000rpm离心l0min，除去油脂和沉淀，收集上清液，即为腹水单克隆抗体。采用硫酸按沉淀和透析法纯化腹水单克隆抗体。
对腹水单克隆抗体进行ELISA效价测定、分子质量鉴定和浓度测定：
1)ELISA效价的测定：采用间接ELISA测定纯化的腹水单克隆抗体，1ug/ml的抗原100u1/孔包被96孔板做ELISA，结果显示纯化后效价为l:210000。
2)抗体的分子质量鉴定：采用SDS-PAGE进行测定单克隆抗体，显示单克隆抗体的重链约为46KD，轻链约为25KD。
3)单克隆抗体浓度测定:紫外分光法测定单克隆抗体在280nm和260nm处的吸光度值A280和A260。结果为蛋白质含量为21mg/ml。
实施例3：病原的临床诊断
1.试剂盒制备
试剂盒采用化学发光免疫法。
1)将兔源抗δ-catenin检测抗体(EliOnco美国)包被在已经活化的磁珠上。取100ul已活化的Mag COOH磁珠，加入100ug兔源抗δ-catenin检测抗体，25℃垂直混合120分钟，磁分离清洗3次，除去游离抗体。再加300ul 1％BSA封闭，25℃垂直混合60分钟，磁分离清洗3次，重新悬浮于1ml PBST溶液中；
2)将吖啶酯标记在抗δ-catenin单克隆抗体LCZ9H4上。标记缓冲液为CB体系，pH为19.5，投料比(摩尔比)为20：1(吖啶/抗体)，终止反应时加入赖氨酸(赖氨酸/吖啶比例为140:1)。标记抗体纯化采用sephadex G-50葡聚糖凝胶柱分离；
3)在各反应孔中，加入50ul上述偶联磁珠，再各孔对应加入待测标准品或者尿液50ul。37℃恒温箱孵育30分钟，磁分离洗板3次。加入吖啶酯标记的抗δ-catenin单克隆抗体LCZ9H4，37℃恒温箱孵育30分钟，磁分离洗板3次后，用化学发光检测仪进行测定。仪器参数：化学发光底物A液和B液各加液50ul，整合时间3S。根据不同浓度的标准品化学发光值绘出标准曲线，将病人的数值与阳性标准孔数值比较，从而得出被测试者尿样本的实际肿瘤标记物δ-catenin浓度。
2.结果检测
用上述试剂盒检测前列腺癌症和正常人尿样本。根据临床样本检测真阳性a，假阳性b，假阴性c，真阴性d，求出检测灵敏度，特异度，总符合率。
表1 患者和正常人群中肿瘤标记物δ-catenin含量(ng/ml)比较表
研究对象分组 病例数 平均值 标准偏差 中位数
患者 100 8.47 2.144 5.47
正常对照 100 0.79 0.287 0.58
*Note：Z＝P＜0.05
由表1可知，癌症患者尿液中肿瘤标记物δ-catenin含量与正常人相比较明显升高，统计学有显著性差异。
对上述200例样本进行临床金标准和本试剂盒的两种检测。计算肿瘤标记物δ-catenin检测试剂盒临床检测的灵敏度、特异性、总符合率、阳性预期值、阴性预期值等统计学结果，结果如表2。
表2 临床金标准和试剂盒δ-catenin检测临床数据归纳表
Figure PCTCN2019129882-appb-000001
灵敏度＝a/(a+c)×100％＝89/100×100％＝89.0％；
特异度＝d/(b+d)×100％＝92/100×100％＝92.0％；
总符合率＝(a+d)/(a+b+c+d)＝(89+92)/200×100％＝90.5％。
由表2可知，该细胞分泌的抗人前列腺小体外泄蛋白抗体对前列腺小体外泄蛋白检测的灵敏度和特异度达到89％以上。
以上所述仅为本发明的实施例，并非因此限制本发明的专利范围，凡是利用本发明说明书内容所作的等效结构变换，或直接或间接运用在其他相关的技术领域，均同理包括在本发明的专利保护范围内。

Claims (9)

  1. 一种杂交瘤细胞LCZ9H4，其特征在于：所述杂交瘤细胞LCZ9H4保藏于中国典型培养物保藏中心，保藏编号为CCTCC No：C2019145。
  2. 一种单克隆抗体，其特征在于：其由权利要求1所述的杂交瘤细胞LCZ9H4分泌产生。
  3. 一种试剂盒，其特征在于：所述试剂盒包括权利要求1所述的杂交瘤细胞或权利要求2所述的单克隆抗体。
  4. 根据权利要求3所述的试剂盒，其特征在于：所述试剂盒为化学发光试剂盒。
  5. 权利要求1所述的杂交瘤细胞或权利要求2所述的单克隆抗体在制备试剂盒中的用途。
  6. 根据权利要求5所述的用途，其特征在于：所述试剂盒是基于免疫检测的试剂盒。
  7. 根据权利要求6所述的用途，其特征在于：所述试剂盒为化学发光试剂盒。
  8. 根据权利要求5所述的用途，其特征在于：所述试剂盒用于检测人前列腺癌。
  9. 权利要求1所述的杂交瘤细胞或权利要求2所述的单克隆抗体在制备用于人前列腺癌诊断的试剂盒中的用途。
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