CN116903739B - 一种抗s100b蛋白的抗体及其应用 - Google Patents
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Abstract
本发明公开了一种抗S100B蛋白的抗体及其应用。本发明所述的抗体包含重链可变区和轻链可变区,所述轻链可变区CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4所示;所述重链可变区CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8所示。
Description
技术领域
本发明涉及生物医学技术领域,具体涉及一种抗人S100B蛋白的抗体及其应用。
背景技术
S100钙结合蛋B(S100B)是一种相对分子质量较小的钙离子结合蛋白,其特征在于它能溶解于100%饱和硫酸铵饱和溶液中,从而得以命名。主要存在于星形胶质细胞和中枢神经系统的胶质细胞中,被认为是神经胶质的标志蛋白。研究表明,在低浓度下S100B具有神经营养作用,在高浓度下可刺激促炎症细胞因子的表达,引起细胞凋亡,发挥其神经毒性作用,促进神经退行性疾病和神经炎症的发展。
S100B蛋白作为特异性血清生化标记物,对脑损伤、神经退行性疾病、脑血管疾病以及恶性黑色素瘤等都有极高的敏感性和特异性。例如,在急性脑梗死患者中血清S100B蛋白水平升高,可反映脑梗死的严重程度,并可预测患者预后;血清S100B蛋白水平也可以反应TBI的严重程度、病情的动态变化及患者的预后;S100B蛋白含量的动态变化可以作为神经系统疾病早期诊断、综合治疗、药物疗效评价及疾病转归预测的重要指标之一。因此,S100B蛋白浓度的检测将为临床诊断和治疗提供重要的判断依据。
S100B具有神经营养作用,而过多的S100B具有神经毒性作用。S100B通过一系列细胞转导途径促进细胞凋亡,参与炎症反应,从而参与多种疾病的病理生理,在神经系统多种疾病中具有广阔研究前景。另外,S100B也可作为一种细胞因子参与疾病的病理过程。例如,其在肌细胞增殖中具有重要作用,而肌卫星细胞的增殖和凋亡的减少是肌营养不良症在细胞治疗中成功的关键。鉴于此,S100B蛋白还可作为疾病治疗的新靶点。
发明内容
本发明的目的在于提供一种抗人S100B蛋白的抗体。
本发明的另一目的在于提供编码所述的抗人S100B蛋白的抗体的载体或细胞。
本发明的另一目的在于提供一种检测S100B蛋白的试剂盒。
本发明的另一目的在于提供包含所述的抗人S100B蛋白的抗体的药物组合物。
为实现上述目的,第一方面,本发明提供了一种针对S100B蛋白的抗体或其抗原结合片段,所述抗体或其抗原结合片段包含重链可变区和轻链可变区,
所述轻链可变区CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4所示;
所述重链可变区CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8所示。
所述的S100B蛋白是一种钙结合蛋白,S指英文Soluble可溶的,100是指其在饱和硫酸铵溶液中的溶解度为100%。在本发明中,所述的S100B蛋白是人S100B蛋白。
进一步,所述的抗体或其抗原结合片段的轻链可变区氨基酸序列如SEQ ID NO:1所示,或该序列经替换、缺失或添加一个或几个氨基酸形成的具有同等功能的氨基酸序列;和/或
所述的抗体或其抗原结合片段的重链可变区氨基酸序列如SEQ ID NO:5所示,或该序列经替换、缺失或添加一个或几个氨基酸形成的具有同等功能的氨基酸序列。
如本文中在诸如“A和/或B”的短语中使用的术语“和/或”旨在包括A和B两者;A或B;A(单独);以及B(单独)。同样地,在诸如“A、B和/或C”的短语中使用的术语“和/或”旨在涵盖以下实施方案的每一个:A、B和C;A、B或C;A或C;A或B;B或C;A和C;A和B;B和C;A(单独);B(单独);以及C(单独)。
如本文所用,术语“抗体”是指免疫球蛋白分子,其通过至少一个抗原结合位点识别并特异性结合靶标,诸如蛋白质、多肽、肽、碳水化合物、多核苷酸、脂质或前述的组合。如本文所用,该术语涵盖完整的多克隆抗体、完整的单克隆抗体、单链抗体、抗体片段(诸如Fab、Fab′、F(ab')2和Fv片段)、单链Fv(scFv)抗体、多特异性抗体(诸如双特异性抗体)、单特异性抗体、单价抗体、嵌合抗体、人源化抗体、人抗体、包含抗体的抗原结合位点的融合蛋白,以及包含抗原结合位点的任何其他修饰的免疫球蛋白分子,只要该抗体表现出所需的生物结合活性。抗体可以是五种主要类别的免疫球蛋白中的任一种:IgA、IgD、IgE、IgG和IgM,或其亚类(同种型)(例如IgG1、IgG2、IgG3、IgG4、IgA1和IgA2)。不同类别的免疫球蛋白具有不同的和熟知的亚单位结构和三维构型。抗体可以是裸露的或与其他分子缀合,包括但不限于毒素和放射性同位素。
在本发明的具体实施例中,所述的抗体为IgG2b型。
在本发明的具体实施例中,所述的抗体为单克隆抗体。
在本发明的具体实施例中,所述的抗体或其抗原结合片段由保藏号为CGMCCNO.45492的杂交瘤细胞产生。
第二方面,本发明提供了一种分离的核酸分子,所述核酸分子编码本发明第一方面所述的抗体或其抗原结合片段。
核酸是脱氧核糖核苷酸或核糖核苷酸及其单链、双链或三螺旋形式的聚合物。除非明确限制,否则核酸可以包含与参考天然核酸具有相似的性质的天然核苷酸的已知类似物。核酸包括基因、cDNAs、mRNAs和cRNAs。核酸可以进行合成,或可以源于任何生物来源,包括任何生物。
本发明的核酸可以进行克隆、合成、改变、突变或其组合。用于分离核酸的标准重组DNA和分子克隆技术是本领域已知的。产生碱基对改变、缺失或小插入的定点诱变也是本领域已知的。参见例如,Sambrook等人(编辑)(1989)Molecular Cloning:A LaboratoryManual.Cold SpringHarbor Laboratory Press,Cold Spring Harbor,New York;Silhavy等人(1984)Experiments with Gene Fusions.Cold Spring Harbor LaboratoryPress,Cold Spring Harbor,New York;Glover&Hames(1995)DNACloning:A PracticalApproach,第2版,IRL Press at Oxford UniversityPress,Oxford/New York;Ausubel(编辑)(1995)Short Protocols inMolecular Biology,第3版,Wiley,New York。
第三方面,本发明提供了一种载体,所述的载体含有本发明第二方面所述的核酸分子。
在本发明的具体实施方案中,所述载体优选为表达载体,除了前面所述的核酸分子之外,表达载体还包括与所述核酸分子序列操作性相连的表达调控序列。
表达载体是指可将编码某蛋白的多聚核苷酸插入其中并使蛋白获得表达的一种核酸运载工具。载体可通过转化、转导或转染宿主细胞,使其携带的遗传物质元件在宿主细胞内得以表达。载体的种类包括本领域熟知的细菌质粒、噬菌体、酵母质粒、植物细胞病毒、哺乳动物细胞病毒如腺病毒、逆转录病毒或其他载体。总之,只要能在宿主体内复制和稳定,任何质粒和载体都可以用。在表达载体中,除了含有复制起点外,还可含有标记基因和其他翻译控制元件。
第四方面,本发明提供了一种宿主细胞,所述的宿主细胞含有本发明第二方面所述的核酸分子或本发明第三方面所述的载体。
进一步,所述的宿主细胞包括真核细胞、原核细胞。
所述真核细胞和原核细胞能够用作用于表达本发明第一方面所述抗体或其抗原结合片段的宿主细胞,此类宿主细胞是此类宿主细胞是本领域中众所周知的,并且许多可从美国典型培养物保藏中心(American Type Culture Collection,ATCC)获得。这些宿主细胞尤其包含中国仓鼠卵巢(CHO)细胞、NS0、SP2细胞、海拉细胞(HeLa cell)、小仓鼠肾(BHK)细胞、猴肾细胞(COS)、人肝细胞癌细胞(例如,Hep G2)、A549细胞、3T3细胞、HEK-293细胞和许多其它细胞系。哺乳动物宿主细胞包含人、小鼠、大鼠、狗、猴、猪、山羊、牛、马和仓鼠细胞。可以使用的其它细胞系是昆虫细胞系(例如,草地贪夜蛾(Spodopterafrugiperda)或粉纹夜蛾(Trichoplusiani))、两栖动物细胞、细菌细胞、植物细胞和真菌细胞。真菌细胞包含酵母和丝状真菌细胞。
在本发明的具体实施例中,所述的宿主细胞为保藏号为CGMCCNO.45492的杂交瘤细胞。
第五方面,本发明提供了一种检测S100B蛋白的试剂盒,所述的试剂盒含有本发明第一方面所述的抗体或其抗原结合片段。
进一步,所述的试剂盒还含有第二抗体和用于检测的酶或荧光或放射标记物,以及缓冲液。
进一步,所述的第二抗体为抗本发明第一方面所述抗体或其抗原结合片段的抗抗体。
第六方面,本发明提供了一种药物组合物,所述的药物组合物包含本发明第一方面所述的抗体或其抗原结合片段。
本发明所述的药物组合物还可以包含药学上可接受的水性载剂。通常将载剂理解为适合于存储、运输、配制和/或施用化合物例如药物活性化合物,特别是根据本公开的抗体的材料。例如,载剂可以是生理上可接受的液体,其适合于存储、运输和/或施用药物活性化合物,特别是根据本公开的抗体。
制备本文所述的药物组合物用于注射或输注给患者。在一些实施方案中,可以制备药物组合物用于静脉内、动脉内或心室内输注。在其他实施方案中,可以制备药物组合物用于静脉内、动脉内、心室内、髓内、腹膜内、鞘内、心室内或皮下注射。在特定实施方案中,制备药物组合物用于肌肉注射。本文所述的药物组合物可以是药学上可接受的无菌水溶液,其表现出合适的pH、等渗性和稳定性,用于施用至人类对象。适合于本文所述的药物组合物的配制的水性载体包括水(例如,无菌水、USP注射用水),以及等渗载体,例如氯化钠注射液、林格注射液、乳酸盐林格注射液。
第七方面,本发明提供了如下任一方面应用,所述应用包括;
(1)本发明第一方面所述的抗体或其抗原结合片段、本发明第二方面所述的核酸分子、本发明第三方面所述的载体、本发明第四方面所述的宿主细胞、或本发明第五方面所述的试剂盒在检测S100B蛋白中的应用;
(2)本发明第一方面所述的抗体或其抗原结合片段、本发明第二方面所述的核酸分子、本发明第三方面所述的载体、本发明第四方面所述的宿主细胞、或本发明第五方面所述的试剂盒在制备诊断S100B蛋白相关疾病中的应用;
(3)本发明第一方面所述的抗体或其抗原结合片段、本发明第二方面所述的核酸分子、本发明第三方面所述的载体、本发明第四方面所述的宿主细胞、或本发明第六方面所述的药物组合物在制备治疗S100B蛋白相关疾病中的应用。
在本发明中,所述的S100B蛋白相关疾病包括脑梗死、脑损伤、黑色素瘤、精神分裂症、卒中后抑郁、神经退行性疾病,优选的,所述的脑损伤包括颅脑损伤、脑出血损伤、脑血管疾病致脑损伤、脑膜炎致脑损伤。
第八方面,本发明提供了一种非诊断目的的检测S100B蛋白方法,所述的方法包括:
提取含有S100B蛋白的样品,将获取的样品与本发明第一方面所述的抗体接触,检测样品与抗体的免疫反应,确定样品中S100B蛋白的表达水平。
与现有技术相比,本发明具有如下有益效果:
本发明首次公开了一种检测S100B蛋白的抗体,经过实验获得的抗体具有很好的特异性和亲和力,可以用于检测人样本中S100B蛋白的浓度。
附图说明
图1为抗S100B单克隆抗体SDS-PAGE电泳分析结果图,图中标号M为Marker,标号1为纯化后的4B3782细胞株。
图2为抗S100B单克隆抗体亚类鉴定结果图,4B3782细胞株为IgG2b型。
图3为抗S100B单克隆抗体Western Blot免疫检测真核表达S100B蛋白的结果图,图中标号1为转染pcDNA3.4-S100B质粒的293细胞裂解液,标号2为转染pcDNA3.4空载体的293细胞裂解液。
具体实施方式
为了对本发明的技术特征、目的和有益效果有更加清楚的理解,现结合具体实施例对本发明的技术方案进行以下详细说明,应理解这些实例仅用于说明本发明而不用于限制本发明的范围。实施例中,各原始试剂材料均可商购获得,未注明具体条件的实验方法为所属领域熟知的常规方法和常规条件,或按照仪器制造商所建议的条件。
生物材料保藏说明
本发明的单克隆抗体杂交瘤细胞株:小鼠杂交瘤细胞株4B3782,保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),保藏中心登记入册编号为CGMCCNo.45492,保藏日期为:2023年03月03日。中国微生物菌种保藏管理委员会普通微生物中心的地址为:北京市朝阳区北辰西路1号院3号,邮编100101。
实施例1:免疫原制备
根据Genebank中公布的人S100B蛋白全长基因序列,由北京擎科生物科技有限公司合成全长基因,并连接入pET-28a质粒中,获得重组质粒pET-28a-S100B。将重组质粒转化入BL21感受态细胞,挑取单菌落于2mL含氨苄西林钠的LB液体培养基中,37℃振荡培养过夜,次日接种于200mL新鲜的LB液体培养基中,37℃、180rpm培养4h至对数生长期,加200μL1mol/L的IPTG诱导液,30℃诱导12-14h。4℃,6000rpm离心10min收集诱导后的菌体;用25mmol/L Tris-HCl(pH8.5)重悬菌体,冰浴超声;4℃,12000rpm离心10min收集上清备用。进行SDS-PAGE凝胶电泳,分析S100B蛋白主要以可溶形式表达在上清液中。将pET-28a-S100B菌液扩大培养,用Ni柱进行亲和层析纯化蛋白,用含25mM咪唑、250mM咪唑的25mMTris-HCl(pH=8.5)溶液洗脱,分别收集蛋白峰,电泳分析纯化产物,取250mM咪唑洗脱蛋白作为免疫原(S100B抗原)。
实施例2:杂交瘤细胞株建立
取纯化后S100B蛋白作为免疫原,采用8周龄BALB/c雌性小鼠,抗原加等量弗氏完全佐剂背部及腹腔注射小鼠(50μg/只);第四周和第八周进行第2次和第3次相同剂量免疫,用弗氏不完全佐剂,3天后取脾细胞进行融合。复苏SP20骨髓瘤细胞,培养至其处于对数生长期。取免疫的BALB/c小鼠,摘除眼球采血供阳性对照血清,同时颈脱位处死小鼠,用75%酒精消毒体表3-5min,取其脾脏,制备脾细胞悬浮液。
取上述脾细胞与骨髓瘤细胞按照5:1的比例,在无血清的1640培养基中混匀,1200rpm离心5分钟,充分吸取上清,轻轻震荡离心管底部,振散细胞,在45-60秒内加入1mL预热过的50%PEG融合细胞,边加边轻轻摇匀,加完后静置90秒,加入无血清的1640培养基终止融合(第一分钟加1mL,第二分钟加2mL,第三分钟加8mL),37℃静置10min,1200rpm离心5分钟,沉淀用HAT培养基悬浮,分装到96孔含有饲养细胞的细胞板中,37℃、5%CO2的细胞培养箱中培养。细胞培养箱中培养5天后,用HAT培养基换液一次,第10天用HAT培养基换液,等到融合细胞覆盖孔底10%-50%时,采用间接ELISA法筛选阳性克隆,保存克隆株,定名为小鼠杂交瘤细胞株4B3782。
实施例3单克隆抗体制备及其亚型分析
4B3782杂交瘤细胞,用含10%胎牛血清的1640培养基培养。每只BALB/c雄性小鼠腹腔注射0.5mL液体石蜡。10天后收集细胞,用10mL生理盐水重悬细胞,每只小鼠腹腔注射0.5mL(细胞密度大约为1×107个/mL)。2周后,收集腹水。以Thermo公司Melon GelMonoclonal IgG Purification Kit试剂盒进行抗体纯化,纯化后的抗体分装后于-20℃保存。
采用Pierce Papid Isotyping Kit-Mouse试剂盒抗体亚型鉴定,首先用样本稀释液将抗体稀释为500ng/mL,然后每孔加入150μl稀释好的抗体,10min后观察记录结果。结果显示4B3782单克隆抗体为小鼠IgG2B亚型,如图2所示。
实施例4:抗人S100B蛋白单克隆抗体可变区序列测定
培养小鼠杂交瘤细胞株4B3782,Trizol法提取杂交瘤细胞总RNA,Thermo Fisher公司High Capacity cDNA Rever Transcription Kit试剂盒逆转录cDNA后,根据《重组抗体》(科学出版社,沈倍奋主编,2005年出版)中鼠单抗引物序列,设计并由北京擎科生物科技有限公司合成该抗体的重轻链引物,PCR扩增(扩增程序为:95℃预热3min,进行30个循环(95℃30秒,60℃30秒,72℃60秒),最后72℃延伸10min),连接PMD18-T载体,转入大肠杆菌JM109中,挑选阳性克隆进行测序。将测定序列在BLAST网站(https://www.ncbi.nlm.nih.gov/igblast/)比对分析小鼠来源单克隆抗体CDR区序列。
经序列分析后发现,4B3782单克隆抗体轻链可变区氨基酸序列为109个氨基酸,其序列如下:
IVLTQSPASLAVSLGQRATISYRASKSVSTSGYSYMHWNQQKPGQPPRLLIYLVSNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHIRELTRSEGGPSWK(SEQ ID NO.1),其中带有下划线的序列依次为CDR1、CDR2和CDR3,其中,CDR1位于26-35aa,氨基酸序列为KSVSTSGYSY(SEQ ID NO.2);CDR2位于53-58aa,氨基酸序列为LVS(SEQ ID NO.3);CDR3位于92-99aa,氨基酸序列为QHIRELTR(SEQ ID NO.4)。
4B3782单克隆抗体重链可变区氨基酸序列为118个氨基酸,其序列如下:EVQLVESGGGLVQPGGSRKLSCAASGFTFSRFGMHWVRQAPEKGLEWVAYISSGGGTIYYADTVKGRFTVSRDNPKNTLFLQMTSLRSEDTAMYYCARDYGSDYFDYWGQGTTLTVSS(SEQ ID NO.5),其中带有下划线的序列依次为CDR1、CDR2和CDR3,其中,CDR1位于26-33aa,氨基酸序列为GFTFSRFG(SEQ ID NO.6);CDR2位于51-58aa,氨基酸序列为ISSGGGTI(SEQ ID NO.7);CDR3位于97-107aa,氨基酸序列为ARDYGSDYFDY(SEQ ID NO.8)。
实施例6:抗人S100B蛋白单克隆抗体活性测定
1、酶联免疫法
用碳酸盐包被缓冲液将S100B蛋白稀释成浓度为2.5μg/mL,每孔包被100μl,4℃过夜;用洗液洗板2次,每孔300μl;每孔加入120μl封闭液室温封闭6小时;用洗液洗板5次,每孔300μl;用抗体稀释液将4B3782单克隆抗体进行梯度稀释,稀释比例为1:3000、1:9000、1:27000、1:81000、1:243000、1:729000和1:2187000,以每孔100μl加样并设置空白孔(100μl抗体稀释液);37℃孵育30min;用洗液洗板5次,每孔300μl;HRP标记的山羊抗小鼠二抗37℃孵育20min;用洗液洗板5次,每孔200μl;加新鲜配制的底物溶液,每孔100μl,37℃孵育10分钟;每孔加入50μl 2M H2SO4终止反应,采用酶标仪波长450nm测定各孔的吸光值,结果如表2所示,所制备的单克隆抗体可以特异性识别人S100B蛋白,并且所制备的单克隆抗体的效价大于1:243000,表明所制备的单克隆抗体对S100B蛋白具有非常高的亲和力。
表1小鼠抗人S100B蛋白单克隆抗体的效价测定(OD值)
稀释比例 | 1:3000 | 1:9000 | 1:27000 | 1:81000 | 1:243000 | 1:729000 | 1:2187000 |
OD450 | 3.123 | 2.9985 | 2.767 | 1.8245 | 0.945 | 0.374 | 0.1345 |
2、Western Blot免疫印迹法
将HEK293T细胞按每孔5×105的比例接种于6孔板中,铺制2个孔,次日,待细胞生长至80%左右(约12-16h)时,在每个孔中采用jetPRIME试剂将1μgpcDNA3.4/S100B质粒和空载体转染HEK293T细胞。培养24h后收集细胞,预冷的PBS洗2次,加入Thermo公司M-PERMammalian Protein Extraction Reagent裂解液冰上裂解细胞5min,4℃16200×g离心15min,收集上清,加入等体积的2×SDS上样缓冲液,沸水浴煮样5min后,10%SDS-PAGE,转印至PVDF膜,5%脱脂奶粉室温封闭1h,加入浓度为2μg/ml的4B3782单克隆抗体,4℃孵育过夜;0.1%TBST洗3次后,加入羊抗鼠-HRP抗体,室温孵育1h,0.1%TBST洗3次后,ECL化学发光液显色,保存结果,结果如图3所示,说明制备的抗体能特异性识别外源的S100B蛋白。
3、双抗体夹心法检测S100B抗原
用碳酸盐包被缓冲液将4B3782单克隆抗体稀释成浓度为5μg/mL,每孔包被100μl,4℃过夜;用洗液洗板2次,每孔300μl;每孔加入120μl封闭液室温封闭6小时;每孔加入S100B全长蛋白100μl,浓度分别为500、250、125、62.5和0ng/ml,37℃孵育30min;用洗液洗板5次,每孔300μl;加入HRP标记的4B3782抗体作为检测抗体,37℃孵育30min;洗液洗板5次,加入TMB底物,37℃显色10min;加入终止液50μl,采用酶标仪波长450nm测定各孔的吸光值。
表2双抗体夹心法检测S100B抗原实验结果
抗原浓度ng/ml | 500 | 250 | 125 | 62.5 | 0 |
OD450 | 2.670 | 2.338 | 1.695 | 1.250 | 0.102 |
需要说明的是:以上实施例仅用于说明本发明的实施过程和特点,而非限制本发明的技术方案,尽管参照上述实施例对本发明进行了详细说明,本领域的普通技术人员应当理解:依然可以对本发明进行修改或者等同替换,而不脱离本发明的精神和范围的任何修改或局部替换,均应涵盖在本发明的保护范围当中。
Claims (12)
1.一种针对S100B蛋白的抗体或其抗原结合片段,其特征在于,所述抗体或其抗原结合片段包含重链可变区和轻链可变区,
所述轻链可变区CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:2、SEQ ID NO:3、SEQID NO:4所示;
所述重链可变区CDR1、CDR2、CDR3的氨基酸序列分别如SEQ ID NO:6、SEQ ID NO:7、SEQID NO:8所示。
2.根据权利要求1所述的抗体,其特征在于,所述的抗体或其抗原结合片段的轻链可变区氨基酸序列如SEQ ID NO:1所示,或该序列经替换、缺失或添加一个或几个氨基酸形成的具有同等功能的氨基酸序列;和/或
所述的抗体或其抗原结合片段的重链可变区氨基酸序列如SEQ ID NO:5所示,或该序列经替换、缺失或添加一个或几个氨基酸形成的具有同等功能的氨基酸序列。
3.一种分离的核酸分子,其特征在于,所述核酸分子编码权利要求1或2所述的抗体或其抗原结合片段。
4.一种载体,其特征在于,所述的载体含有权利要求3所述的核酸分子。
5.一种宿主细胞,其特征在于,所述的宿主细胞含有权利要求3所述的核酸分子或权利要求4所述的载体。
6.根据权利要求5所述的宿主细胞,其特征在于,所述的宿主细胞为保藏号为CGMCCNO.45492的杂交瘤细胞。
7.一种检测S100B蛋白的试剂盒,其特征在于,所述的试剂盒含有权利要求1或2所述的抗体或其抗原结合片段。
8.根据权利要求7所述的试剂盒,其特征在于,所述的试剂盒还含有第二抗体和用于检测的酶或荧光或放射标记物,以及缓冲液。
9.根据权利要求8所述的试剂盒,其特征在于,所述的第二抗体为抗权利要求1或2所述抗体或其抗原结合片段的抗体。
10.一种药物组合物,其特征在于,所述的药物组合物包含权利要求1或2所述的抗体或其抗原结合片段。
11.如下任一方面应用,其特征在于,所述应用包括;
(1)权利要求1或2所述的抗体或其抗原结合片段、权利要求3所述的核酸分子、权利要求4所述的载体、权利要求5或6所述的宿主细胞、或权利要求7-9任一项所述的试剂盒在制备检测S100B蛋白的产品中的应用;
(2)权利要求1或2所述的抗体或其抗原结合片段、权利要求3所述的核酸分子、权利要求4所述的载体、权利要求5或6所述的宿主细胞、或权利要求7-9任一项所述的试剂盒在制备诊断S100B蛋白相关疾病的产品中的应用。
12.一种非诊断目的的检测S100B蛋白方法,其特征在于,所述的方法包括:
提取含有S100B蛋白的样品,将获取的样品与权利要求1或2所述的抗体或其抗原结合片段接触,检测样品与抗体的免疫反应,确定样品中S100B蛋白的表达水平。
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