CN114994308A - 一种检测抗Desmoglein 1-IgG抗体的试剂盒 - Google Patents
一种检测抗Desmoglein 1-IgG抗体的试剂盒 Download PDFInfo
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Abstract
本发明提供了一种检测抗Desmoglein 1‑IgG抗体的试剂盒,涉及生物检测技术领域。本发明提供了一种检测抗人Desmoglein 1自身抗体的试剂在制备肾病综合征检测工具中的应用,并基于此建立了检测抗Desmoglein 1‑IgG抗体的试剂盒,所述试剂盒基于靶点Desmoglein 1及其相应自身抗体。本发明所述试剂盒可通过与抗原蛋白Desmoglein 1的免疫反应对来自血液、血清、组织或体液中的自身抗体进行检测。本发明试剂盒既可以定性也可以定量检测抗Desmoglein 1‑IgG抗体,能够诊断或辅助诊断血管内皮损伤,填补了国内外鉴别血管内皮损伤自身抗体的空白。
Description
技术领域
本发明属于生物检测技术领域,具体涉及一种检测抗Desmoglein 1-IgG抗体的试剂盒。
背景技术
血液、血管和心脏组成了人体的血液循环系统。血液循环系统中的血液在血管中流动,流经心脏、肺、肝等全身脏器。血管的最里层附着有血管内皮细胞,血管内皮细胞是介于血流和血管壁组织之间的一层单核细胞,可通过自分泌、内分泌、旁分泌三种途径分泌一系列NO、PGI2、ET-1等血管活性物质发挥调节血管紧张性、抗血栓形成、抑制平滑肌细胞增殖及血管壁炎症反应等功能。NO是内皮细胞产生最重要的舒血管因子,由内皮细胞的NO合酶(eNOs)作用于L-精氨酸产生,NO可扩散至血管壁平滑肌细胞激活鸟氨酸环化酶,介导cGMP调控的血管舒张。不仅如此,NO还具有抑制血小板聚集、抑制单核细胞粘附于内皮细胞、抑制平滑肌细胞增殖等作用。然而血管内皮在受到一系列有害因素作用时,内皮细胞释放的舒血管因子减少,缩血管因子增多,打破血管平衡稳态,最终导致一系列心血管事件的发生。血管内皮细胞自身抗体会造成血管内皮细胞损伤,诱发血液循环系统功能障碍,从而导致心脏、肺、肝等脏器的损伤,引发各个脏器相关的疾病,包括肾病综合征。
内皮细胞是血管内侧的单层细胞,具有高代谢活性,在许多生理过程中起关键作用,包括调控血管舒缩张力、血液与组织之间的血细胞运输、维持血液流动性,通透性,血管生成和固有和适应性免疫,参与大多数疾病的病理生理过程,是病理生理学的主要决定因素或受害者。与其余器官相比,肾脏具有最丰富和最多样化的内皮细胞群体,这种广泛的多样性包括肾脏内皮细胞有助于跨肾各个部分的不同转运能力以及不同的内皮细胞承受环境中的氧气含量和渗透压不同。因而,血管内皮细胞损伤可导致包括肾病综合征在内的多种脏器疾病,危害严重。
微小病变病(MCD)是儿童肾病综合征的主要原因,占成人肾病综合征的10~15%。微小病变病患者肾小球在光学显微镜下看起来基本正常,在电子显微镜下可见的唯一组织病理学异常是弥漫性足细胞足突融合消失。因此,MCD被认为是一种原发性足细胞疾病。皮质类固醇治疗后蛋白尿完全缓解是MCD的标志,一般而言进行性肾功能衰竭很少见。然而,MCD会导致严重的并发症。在成人中观察到的与疾病相关的并发症主要包括静脉血栓形成和需要临时透析的严重急性肾损伤。此外,由于MCD的特点是慢性、复发性病程,因此通常需要延长免疫抑制治疗以维持蛋白尿缓解。然而,长期免疫抑制治疗会增加严重感染的风险,并带来恶性肿瘤的长期风险。
尽管观察到的足细胞损伤是MCD的主要经典特征,但疾病机制可能还涉及肾小球血管内皮细胞。尽管如此,至今仍然不清楚造成肾小球内皮细胞损伤的致病因子到底是什么。申请人研究团队通过前期的研究在MCD和原发性局灶节段性肾小球硬化(FSGS)肾病综合征患者体内筛选和鉴定到了一系列的肾小球血管内皮细胞自身抗体。动物实验证实这些肾小球血管内皮细胞自身抗体会引起小鼠肾小球血管内皮细胞严重损伤。体外细胞培养实验也表明这些自身抗体会影响血管内皮细胞的形态和功能。临床研究更是表明这些肾小球血管内皮细胞自身抗体与患者的高凝状态以及不良预后有关。但是目前关于肾小球血管内皮细胞自身抗体与肾病综合征发病机制之间的关联无法直接确定,而且目前市场上缺少抗Desmoglein 1抗体是的临床检测试剂盒。
发明内容
有鉴于此,本发明的目的在于提供一种检测抗Desmoglein 1-IgG抗体的试剂盒,所述试剂盒既可以定性也可以定量检测抗Desmoglein 1-IgG抗体,具有良好的准确性、灵敏度、特异性及检测速度。
为了实现上述发明目的,本发明提供以下技术方案:
本发明提供了一种检测抗人Desmoglein 1自身抗体的试剂在制备血管内皮损伤检测工具中的应用。
优选的,所述检测工具包括检测试剂盒。
优选的,所述抗人Desmoglein 1自身抗体包括抗人Desmoglein 1-IgG抗体。
本发明还提供了一种检测抗Desmoglein 1-IgG抗体的试剂盒,包括独立包装的抗原蛋白Desmoglein 1、固相载体和标记抗体;
所述抗原蛋白Desmoglein 1包括蛋白登录号为BC153001的整体或片段。
优选的,所述标记抗体包括利用酶标记、荧光编码微球、磁条芯片、化学发光剂标记或生物素标记的二抗;
所述二抗包括抗人IgG抗体。
优选的,所述固相载体包括硝酸纤维素膜、磁微粒和/或酶标微孔板。
优选的,在所述抗原蛋白Desmoglein 1的N端或C端还结合有标签,所述标签包括His标签、硫氧还蛋白、GST标签、麦芽糖结合蛋白、谷胱甘肽转移酶的SA标签、c-Myc标签、Flag标签或生物素标签。
优选的,所述试剂盒的检测样本包括血液、血清、组织或体液。
有益效果:本发明提供了一种检测抗人Desmoglein 1自身抗体的试剂在制备血管内皮损伤检测工具中的应用,并基于此建立了检测抗Desmoglein1-IgG抗体的试剂盒,所述试剂盒基于靶点Desmoglein 1及其相应自身抗体。本发明所述试剂盒可通过与抗原蛋白Desmoglein 1的免疫反应对来自组织或体液中的自身抗体进行检测;本发明首次鉴定了针对Desmoglein 1的IgG自身抗体,利用本发明试剂盒检测298例肾病综合征患者血清中抗Desmoglein 1-IgG抗体,结果显示有120例患者抗Desmoglein 1-IgG抗体阳性,即抗Desmoglein 1-IgG抗体阳性检出率为40.95%,本发明所述血管内皮细胞自身抗体的检测,能够实现肾病综合征的有效诊断,为研究肾病综合征的分子机制和临床诊疗提供依据。且本发明试剂盒利用人抗标签肽的IgG抗体作为标准品以及结合生物素-亲和素放大系统、磁微粒化学发光免疫分析大大提高了检测的准确性、灵敏度、特异性及检测速度。本发明试剂盒既可以定性也可以定量检测抗Desmoglein 1-IgG抗体,为研究肾病综合征的分子机制和临床诊疗提供依据。
附图说明
图1为肾小球血管内皮细胞上的Desmoglein 1蛋白是肾病综合征病人体内自身抗体针对的主要靶抗原;其中A:一抗为健康人血清的二维电泳蛋白点;B:一抗为肾病综合征患者血清的二维电泳蛋白点;C:靶抗原Desmoglein1蛋白的质谱鉴定;
图2为表达的重组蛋白Desmoglein 1的SDS-PAGE鉴定图;
图3为固相膜免疫试剂盒检测肾病综合征患者血清中抗Desmoglein1-IgG抗体;
图4为磁微粒化学发光免疫分析试剂盒检测抗Desmoglein 1-IgG抗体的原理示意图;
图5为抗原蛋白Desmoglein 1包被羧基磁微粒示意图;
图6为各类肾病病人中抗Desmoglein 1-IgG抗体的检测情况,其中NS:肾病综合征,HP:过敏性紫癜,HPN:紫癜性肾炎,KD:川崎病,NC:健康儿童;
图7为抗Desmoglein 1-IgG抗体与肾小球血管内皮细胞损伤标志物线性相关。
具体实施方式
本发明提供了一种检测抗人Desmoglein 1自身抗体的试剂在制备血管内皮损伤检测工具中的应用。
血液、血管和心脏组成了人体的血液循环系统,血液循环系统中的血液在血管中流动,流经心脏、肺、肝等全身脏器。血管最里层附着着血管内皮细胞,血管内皮细胞自身抗体会造成血管内皮细胞损伤,诱发血液循环系统功能障碍,进而导致心脏、肺和肝等脏器的损伤,引发包括肾病综合征在内的各个脏器相关疾病,而不同脏器的血管内皮细胞都是一样的。因而,基于血液循环系统中检测血管内皮细胞自身抗体,可以用于临床血管内皮细胞损伤。本发明所述抗人Desmoglein 1自身抗体的抗原蛋白Desmoglein 1为桥粒芯蛋白1,简称DSG1。在本发明中,所述DSG1优选包括蛋白登录号为BC153001的整体或片段,实施例中为方便说明,以SEQ ID NO.1所示的氨基酸序列(MKKLADISLGKESYPDLDPSWPP QSTEPVCLPQETEPVVSGHPPISPHFGTTTVISESTYPSGPGVLHPKPILDPLGYGNVTVTESYTTSDTLKPSVHVHDNRPASNVVVTERVVGPISGADLHGMLEMPDLRDGSNVIVTERVIAPSSSLPTSLTIHHPRESSNVVVTERVIQPTSGMIGSLSMHPELANAHNVIVTERVVSGAGVTGISGTTGISGGIGSSGLVGTSMGAGSGALSGAGISGGGIGLSSLGGTASIGHMRSSSDHHFNQTIGSASPSTARSRITKYSTVQYSKHHHHHH)为例进行说明,但是不能仅将其认定为本发明的全部保护范围。
本发明对所述抗原蛋白的来源并没有特殊限定,可表达于细菌如大肠杆菌、酵母、哺乳动物细胞中,并经Ni柱亲和层析、分子筛层析、离子交换层析、疏水柱纯化得到。本发明所述抗原蛋白优选包括融合蛋白,如在所述抗原蛋白的N-末端或C-末端结合标签,使其具有某些生物学或物理功能,并特异性结合配体的序列或结构域;所述标签优选包括His标签,硫氧还蛋白,GST标签,麦芽糖结合蛋白,谷胱甘肽转移酶的SA标签,c-Myc标签,Flag标签或生物素标签,所述标签的存在有利于抗原蛋白纯化,固定,沉淀。
本发明所述检测工具优选包括检测试剂盒。本发明所述试剂优选包括检测抗Desmoglein 1-IgG抗体的试剂。
本发明还提供了一种检测抗Desmoglein 1-IgG抗体的试剂盒,包括独立包装的抗原蛋白Desmoglein 1、固相载体和标记抗体;
所述抗原蛋白Desmoglein 1包括蛋白登录号为BC153001的整体或片段。
本发明所述抗原蛋白Desmoglein 1优选与上述相同,在此不再赘述。本发明所述抗原蛋白Desmoglein 1能够固定于固相载体上,所述固相载体优选包括硝酸纤维素膜(NC膜)、荧光编码微球、磁条芯片、磁微粒和/或酶标微孔板。在本发明中,当固相载体的选择不同时,可获得不同的试剂盒,如以磁微粒为固相载体时,可构建检测抗Desmoglein 1-IgG抗体的磁微粒化学发光免疫分析试剂盒;以硝酸纤维素膜为固相载体时,可构建检测抗Desmoglein 1-IgG抗体的固相膜免疫试剂盒,当然也可在同一个试剂盒中共同存在多种不同的固相载体,从而完成定性和定量检测。
本发明所述固定,优选指与Desmoglein 1抗原蛋白不溶于水的固相载体结合,该固相载体或支持物不溶于水,更优选地通过共价键合,静电相互作用,疏水相互作用、或通过二硫键相互作用,最优选通过一个或多个共价键。本发明对所述固定的方法并没有特殊限定,可以是直接固定方式,也可以是间接固定。本发明所述直接固定优选包括通过过滤,离心或层析,将固定化的分子与不溶性载体一起从水溶液中分离出来;还包括可逆或不可逆的方式固定Desmoglein 1抗原蛋白。例如,抗原蛋白通过可裂解的共价键(如可添加含硫醇的试剂来裂解的二硫键)固定于载体,这种固定是可逆的。另外,如抗原蛋白通过在水性溶液中不会裂解的共价键(通过环氧化物基团与将赖氨酸侧链偶联至亲和柱的胺基团的反应形成的键)固定于载体,则固定是不可逆的。本发明所述间接固定,固定对所述抗原蛋白具有特异性亲和力的抗体,然后形成抗原蛋白-抗体复合物以达到固定的效目的。在本发明中,所述固定,针对不同的固相载体,固定方式略有不同,如抗原蛋白Desmoglein 1通过物理吸附方式或非共价键结合到硝酸纤维素膜或聚苯乙烯微孔板上;带有羧基功能团的磁微粒与抗原蛋白Desmoglein 1的氨基结合,抗原蛋白Desmoglein 1通过化学偶联方式结合在磁微粒上。
本发明所述标记抗体优选包括利用酶标记、化学发光剂标记或生物素标记的二抗;所述二抗包括抗人IgG抗体。本发明所述酶标记优选包括辣根过氧化物酶(HorseradishPeroxidase,HRP)标记,所述化学发光剂标记优选包括吖啶酯标记。
本发明所述试剂盒优选还包括抗原稀释液、样品稀释缓冲液、抗原稀释液、底物显色液、洗涤液、标准品、阳性质控品和阴性质控品。本发明所述标准品和阳性质控品优选均为重组人抗标签肽免疫球蛋白G或其片段、或从病人血清中提取抗Desmoglein 1-IgG抗体;所述阴性质控品优选为健康体检者血清。本发明所述底物显色液优选包括TMB、过氧化氢、AMPPD、4-MUP和BCIP中的任意一种;所述抗原稀释液优选为含有0.15mol/L NaCl、1%(W/V)TritonX-100的1×PBS,pH7.35;所述样品稀释缓冲液优选为含有10%(W/V)BSA的0.01MPBS,pH7.35;所述抗体稀释液优选为含有1M D-glucose、2%(W/V)甘油、0.3%(W/V)Tween20的0.01M PBS,pH7.35;所述洗涤液优选为含有0.15mol/LNaCl、10%(W/V)甘油、1%(W/V)TritonX-100的1×PBS,pH7.35。
本发明所述检测试剂盒,利用间接法反应原理,首先将Desmoglein 1抗原吸附于固相载体作为包被抗原,然后加入阳性质控品或标准品或待检样本进行孵育,再加入标记二抗反应后,若待检血清中含有抗Desmoglein 1-IgG抗体,则形成包被抗原Desmoglein 1-待检血清抗Desmoglein 1-IgG抗体-标记抗人IgG抗体三元复合物,最后利用光显色法、化学发光法、荧光发光法来检测光信号,以达到定性或定量分析人血清中抗Desmoglein 1-IgG抗体的目的。本发明所述检测样本优选包括血液、血清、组织或体液。本发明所述组织优选包括肾活检组织;所述体液优选为本领域常规意义上的体液,包括细胞内液和细胞外液两类,其中细胞外液尤以血浆为主。
本发明所述检测试剂盒,涉及固相膜免疫定性分析人血清中抗Desmoglein 1-IgG抗体,以人抗标签肽的IgG抗体作为标准品,大大提高了检测准确性。固相膜免疫定性检测操作简单,试剂用量较少,比传统ELISA节约近10倍;另外NC膜吸附能力极强接近100%,微量抗原能够完全吸附固定在NC膜上;吸附抗原或抗体或已有结果的NC膜可长期保存(-20℃可保存半年),且不影响其活性;另外本发明固相膜免疫定性检测人血清中抗Desmoglein1-IgG抗体的试剂盒引入生物素-亲和素放大系统,大大提高了检测灵敏度。本发明所述检测试剂盒中还涉及磁微粒化学发光免疫分析定量检测人血清中抗Desmoglein 1-IgG抗体,利用磁微粒为固相载体,其直径仅为1.0μm,大大增加了包被表面积,增加了抗原的吸附量,提高了反应速度,也使清洗分离更简便,从而减少污染,降低交叉感染概率。另一方面,采用吖啶酯发光剂直接标记抗人IgG,其化学反应简单、快速、无须催化剂;吖啶酯化学发光为闪光型,其通过起动发光试剂(H2O2、NaOH)0.4s后发射强度即可达到最大,半衰期为0.9s,2s内基本结束,便于快速检测。
下面结合实施例对本发明提供的一种检测抗Desmoglein 1-IgG抗体的试剂盒进行详细的说明,但是不能把它们理解为对本发明保护范围的限定。
实施例1
血管内皮细胞上的Desmoglein 1蛋白是肾病综合征病人体内自身抗体针对的主要靶抗原
(1)血管内皮细胞总蛋白的提取:培养血管内皮细胞株(EAhy926),用PBS洗涤2~3次,然后用聚焦超声仪(Covaris S220,Gene)在含有30mM Tris-HCl、8M尿素、4%CHAPS和蛋白酶抑制剂(#ab65621;Abcam,1:200稀释)的裂解缓冲液中冰上进行充分裂解,然后将样本置于离心机,12000g,4℃,离心30min。收集上清,即为收集到的血管内皮细胞总蛋白。利用BCA蛋白浓度测定试剂盒测定收集到的血管内皮细胞总蛋白浓度。
(2)二维电泳:提取血管内皮细胞总蛋白进行二维电泳后转到硝酸纤维素膜上,分别用健康人和肾病综合征病人的血清作为一抗进行孵育,之后加上二抗进行显影,见图1中A和图1中B。
(3)基质辅助激光解吸/电离飞行时间质谱分析:将步骤(2)显影后进行阳性点的差异分析,选取二维电泳胶上肾病综合征病人强阳性,而健康人阴性或者弱阳性的蛋白点,从凝胶上取下选中的蛋白点,将干燥后的凝胶用胰蛋白酶(0.1μg/μL)进行消化,然后向反应混合物中加入10μL的25mM碳酸氢铵,37℃孵育过夜,然后用三氟乙酸(0.1%)从凝胶中提取肽。用基质辅助激光解吸/电离飞行时间质谱分析(MALDI-TOF-MS)质谱仪对提取的肽进行分析,得到肽质量图谱,鉴定为Desmoglein 1蛋白(图1中C)。
实施例2
重组抗原蛋白Desmoglein 1表达及纯化
利用基因工程的方法以编码Desmoglein 1蛋白的基因为模板,进行PCR扩增,然后构建表达载体进行蛋白表达。本发明表达的抗原蛋白上含有His标签的标签肽。表达的重组蛋白经镍柱亲和层析进行纯化,最后利用SDS-PAGE鉴定重组蛋白Desmoglein 1的分子量为32KDa,见图2。
实施例3
根据抗原Desmoglein 1包被浓度(20μg、40μg、60μg、80μg、100μg五个包被浓度)、各反应时间(10min,20min、30min、40min)和温度(25℃、37℃)、酶标二抗最佳稀释度(1:200、1:400、1:800、1:1600四个稀释度)等4个因素选择正交表,每个因素按2水平重复测定标准阳性血清和标准阴性血清,选择阳性血清的最高光信号值(P)和阴性血清的最低光信号值(N)的比值(P/N)。
通过正交设计,本试剂盒最佳抗原Desmoglein 1包被浓度为60μg/mL、固相膜免疫检测抗Desmoglein 1-IgG抗体试剂盒最佳抗原抗体反应温度为25℃、最佳抗原抗体反应时间20min及最佳生物素标记抗人IgG抗体最佳工作稀释度为1:400;磁微粒化学发光免疫分析检测抗Desmoglein 1-IgG抗体试剂盒最佳抗原抗体反应温度为37℃、最佳抗原抗体反应时间20min及最佳吖啶酯标记抗人IgG抗体最佳工作稀释度为1:400。
实施例4
用于检测抗Desmoglein 1-IgG抗体的固相膜免疫试剂盒的制备
4.1用于检测抗Desmoglein 1-IgG抗体的固相膜免疫试剂盒的组成:
抗原:重组蛋白Desmoglein 1
固相载体:Satourius CN140硝酸纤维素膜
阳性质控品(标准品):人抗His标签免疫球蛋白G(购自湖州英创)
阴性质控品:健康体检者血清
标记抗体:生物素标记抗人IgG抗体
抗原稀释液
样品稀释缓冲液
抗体稀释液
洗涤液
酶工作液:碱性磷酸酶-链霉亲和素
底物显色液:BCIP显色液。
4.2用于检测抗Desmoglein 1-IgG抗体的固相膜免疫试剂盒的检测步骤如下:
4.2.1包被、封闭:将10μL浓度为60μg/mL的Desmoglein 1抗原直接点于硝酸纤维素膜上置37℃孵育箱中干燥20min,将硝酸纤维素膜置于检测板中,加入200μL 5%(W/V)BSA于37℃温盒中封闭20min,弃去封闭液后用洗涤液洗2次;
4.2.2抗原孵育:向检测板内加入10μL用稀释液稀释的抗体标准品或待检血清,同时做阴性对照、阳性对照,25℃孵育20min,每个样品设置3个平行孔;
4.2.3二抗孵育:弃去检测板内液体,洗涤液洗5次×1min,加入20μL1:400生物素标记抗人IgG抗体,25℃孵育20min;
4.2.4显色:弃去检测板内液体,洗涤液洗3次×3min,加500μL碱性磷酸酶-链霉亲和素,室温孵育20min,弃去检测板内液体,洗涤液洗3次×3min,然后加入BCIP显色液,室温反应20min,用流水冲洗检测板,终止酶反应。取出测试硝酸纤维素膜条用吹风机吹干膜条,用比色卡肉眼定性判定,出现明显棕色斑点者为阳性见图3,或将膜条置于显影仪上扫描,显影仪自带的分析软件以参考标准品浓度作为纵坐标、仪器读取的灰度值作为横坐标,绘制标准曲线对血清中抗Desmoglein 1-IgG抗体水平进行半定量分析。
实施例5
用于检测抗Desmoglein 1-IgG抗体的磁微粒化学发光免疫分析试剂盒的制备
5.1抗Desmoglein 1-IgG抗体化学发光检测试剂盒,包括以下组成部分:
(1)吖啶酯标记的抗人IgG;
(2)与Desmoglein 1抗原偶联的羧基磁珠;
(3)化学发光预激发液H2O2和激发液NaOH;
(4)抗Desmoglein 1-IgG抗体系列标准溶液,标准浓度:0μg/mL、4μg/mL、8μg/mL、16μg/mL、20μg/mL、40μg/mL;
(5)抗原稀释液为含有0.15mol/LNaCl、1%(W/V)TritonX-100的1×PBS,pH7.35;所述样品稀释缓冲液为含有10%(W/V)BSA的0.01M PBS,pH7.35;所述抗体稀释液为含有1MD-glucose、2%(W/V)甘油、0.3%(W/V)Tween20的0.01MPBS,pH7.35;所述洗涤液为:含有0.15mol/LNaCl、10%(W/V)甘油、1%(W/V)TritonX-100的1×PBS,pH7.35。
5.2磁珠偶联抗原的制备(图5)
(1)取1mg羧基磁性颗粒于0.5mL离心管中,加入200μL的0.1mol/L MES缓冲液,涡旋混匀,置于磁力架上,静置5min,使磁性颗粒从液体,并丢弃上清液。洗涤3次,然后加入200μL的MES(pH 5.0)缓冲液并涡旋;
(2)加入18μLDesmoglein 1抗原,涡旋,旋转反应管,室温孵育20min;
(3)加入10μL 10mg/mL偶联试剂EDC涡旋,旋转反应管,室温孵育1h;
(4)去除上清液,加入200μL洗涤缓冲液(TBS+0.05%(W/V)Tween-20)洗涤3次;
(5)用含1%(W/V)BSA的缓冲液封闭,重复4次,每次10min。磁性颗粒悬浮液储存在2-8℃。
5.3吖啶酯标记抗体的制备
(1)将100μL抗人IgG抗体放入透析袋中,将透析袋放入1L的标记缓冲液进行透析,期间至少更换3次缓冲液,最后一次透析过夜,标记缓冲液为Na2CO3-NaHCO3缓冲液,pH为10.0,浓度为0.1mol/L;
(2)称取1.7mg吖啶酯NSP-DMAE-NHS溶于447μL无水二甲基甲酰胺DMF中,形成6.5mmol/LNSP-DMAE-NHS DMF溶液;
(3)将透析后的抗体溶液置于500μL离心管中,加入100μL6.5 mmol/L NSP-DMAE-NHS DMF溶液,吖啶酯与抗体的摩尔比为7.4:1,加入200μL标记缓冲液、室温反应45min,加入10μL赖氨酸,继续反应15min终止标记反应;
(4)通过Sephadex G-50柱(1×25cm)将标记物NSP-DMAE-NHS-Ab与游离的NSP-DMAE-NHS分离,用含纯化缓冲液pH为6.3和浓度为0.1mol/L;
(5)分离过程中,用色谱仪检测蛋白质峰,分别测定流出液的化学发光强度和430nm处的吸光度;
(6)收集高光度、高吸光度的洗脱液,加入1%BSA(体积),冰上保存。
5.4样品制备:将样本按1:10比例稀释
5.5用于检测抗Desmoglein 1-IgG抗体的化学发光法试剂盒的检测步骤如下:
(1)100μL待测样品、100μL偶联磁粉悬液、100μL吖啶酯标记二抗依次加入反应管中,摇匀混合,37℃保温20min;
(2)洗涤5次;
(3)充分振摇洗涤后的反应容器,使磁性颗粒均匀分散;
(4)加入100μL化学发光预激发液H2O2,随后加入100μL化学发光激发液NaOH,测定其相对发光强度。样品中抗Desmoglein 1-IgG抗体的含量与其发光强度成正比(图4)。
实施例6
检测血清抗Desmoglein 1-IgG抗体试剂盒的临床应用
6.1受试者纳入从2018年6月到2020年6月诊断出各类肾病的患者,包括298例肾病综合征(NS)、100例过敏性紫癜(HP)、100例紫癜性肾炎(HPN)、100例川崎病(KD)、同时期100例健康儿童(NC)。血清样本取自各类肾病患者和健康对照组。所有受试者在没有进行免疫抑制治疗之前进行第一次血清样本采集。
6.2各类肾病病人中抗Desmoglein 1-IgG抗体的检测情况利用本发明试剂盒检测从2018年6月到2020年6月在诊断出各类肾病的患者血清中抗Desmoglein 1-IgG抗体水平,包括298例肾病综合征、100例过敏性紫癜、100例紫癜性肾炎、100例川崎病及同时期100例健康儿童,结果显示肾病综合征病人中有120名患者抗Desmoglein 1-IgG抗体阳性,而紫癜性肾炎、过敏性紫癜、川崎病以及健康儿童中抗Desmoglein 1-IgG抗体为阴性,见图6。
6.3肾病综合征患者血清抗Desmoglein 1-IgG抗体与血管内皮损伤标志物表达量呈线性相关利用本发明试剂盒检测从2018年6月到2020年6月在诊断出肾病综合征患者血清中抗Desmoglein 1-IgG抗体表达量,并检测患者血清中血管内皮损伤标志物Plvap的表达量,结果显示肾病综合征病人中抗Desmoglein 1-IgG抗体表达量与血管内皮损伤标志物表达量呈线性相关,肾病综合征与血管内皮损伤有关,见图7。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 浙江大学
<120> 一种检测抗Desmoglein 1-IgG抗体的试剂盒
<160> 1
<170> SIPOSequenceListing 1.0
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Claims (8)
1.一种检测抗人Desmoglein 1自身抗体的试剂在制备血管内皮损伤检测工具中的应用。
2.根据权利要求1所述应用,其特征在于,所述检测工具包括检测试剂盒。
3.根据权利要求1所述应用,其特征在于,所述抗人Desmoglein 1自身抗体包括抗人Desmoglein 1-IgG抗体。
4.一种检测抗Desmoglein 1-IgG抗体的试剂盒,其特征在于,包括抗原蛋白Desmoglein 1、固相载体和标记抗体;所述抗原蛋白Desmoglein 1包括蛋白登录号为BC153001的整体或片段。
5.根据权利要求4所述试剂盒,其特征在于,所述标记抗体包括利用酶标记、化学发光剂标记或生物素标记的二抗;所述二抗包括抗人IgG抗体。
6.根据权利要求4所述试剂盒,其特征在于,所述固相载体包括硝酸纤维素膜、荧光编码微球、磁条芯片、磁微粒和/或酶标微孔板。
7.根据权利要求4所述试剂盒,其特征在于,在所述抗原蛋白Desmoglein1的N端或C端还结合有标签,所述标签包括His标签、硫氧还蛋白、GST标签、麦芽糖结合蛋白、谷胱甘肽转移酶的SA标签、c-Myc标签、Flag标签或生物素标签。
8.根据权利要求4所述试剂盒,其特征在于,所述试剂盒的检测样本包括血液、血清、组织或体液。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117074667A (zh) * | 2023-03-02 | 2023-11-17 | 浙江大学 | 多肽或其片段在制备检测血管内皮细胞损伤试剂盒中的应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008097439A2 (en) * | 2007-02-07 | 2008-08-14 | The Trustees Of The University Of Pennsylvania | Anti-autoimmune antibodies for treatment of pemphigus |
US20200123230A1 (en) * | 2018-10-22 | 2020-04-23 | Euroimmun Medizinische Labordiagnostika Ag | Diagnosis of Blistering Autoimmune Disease |
WO2022043415A1 (en) * | 2020-08-27 | 2022-03-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for detecting the presence of pemphigus-specific autoantibodies in a sample |
CN114152754A (zh) * | 2021-11-26 | 2022-03-08 | 钱华 | 一种用于检测人Dsg1 IgG抗体的ELISA试剂盒及其应用 |
-
2022
- 2022-05-07 CN CN202210491000.0A patent/CN114994308A/zh active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008097439A2 (en) * | 2007-02-07 | 2008-08-14 | The Trustees Of The University Of Pennsylvania | Anti-autoimmune antibodies for treatment of pemphigus |
US20200123230A1 (en) * | 2018-10-22 | 2020-04-23 | Euroimmun Medizinische Labordiagnostika Ag | Diagnosis of Blistering Autoimmune Disease |
CN111072768A (zh) * | 2018-10-22 | 2020-04-28 | 欧蒙医学实验诊断股份公司 | 水疱形成性自身免疫疾病的诊断 |
WO2022043415A1 (en) * | 2020-08-27 | 2022-03-03 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for detecting the presence of pemphigus-specific autoantibodies in a sample |
CN114152754A (zh) * | 2021-11-26 | 2022-03-08 | 钱华 | 一种用于检测人Dsg1 IgG抗体的ELISA试剂盒及其应用 |
Non-Patent Citations (1)
Title |
---|
胡智祥主编: "《医院临床检验技术操作规范与实(化)验室管理全书》", 30 August 2004 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117074667A (zh) * | 2023-03-02 | 2023-11-17 | 浙江大学 | 多肽或其片段在制备检测血管内皮细胞损伤试剂盒中的应用 |
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