CN106749659A - 一种抗人crp抗体及其应用 - Google Patents
一种抗人crp抗体及其应用 Download PDFInfo
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- CN106749659A CN106749659A CN201710037823.5A CN201710037823A CN106749659A CN 106749659 A CN106749659 A CN 106749659A CN 201710037823 A CN201710037823 A CN 201710037823A CN 106749659 A CN106749659 A CN 106749659A
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Abstract
本申请涉及抗人C‑反应蛋白(CRP)抗体及其制备方法以及上述抗体在人C‑反应蛋白检测中的应用。本申请发明人制备了多种抗体，并进行配对筛选，获得灵敏度及特异性均能满足需求的抗体组合(P20及P02)；同时其方便大量生产，可满足日后大规模临床应用的需求。对上述抗体组合进行检测体系的调试优化工作，获得操作简便，灵敏度，特异性及相关检测性能均可满足人临床样本检测的人C‑反应蛋白的胶体金免疫层析定量检测卡。
Description
技术领域
本发明属于生物技术领域，具体的涉及两种抗人C-反应蛋白(CRP)抗体及其制备方法以及上述抗体在人C-反应蛋白检测中的应用。
背景技术
C-反应蛋白(C reactive protein，CRP)是蛋白质家族中的一员，是一种急性期反应蛋白，在组织损伤和细菌感染时血浆浓度急剧升高，是机体重要的防御分子，主要由肝脏产生并分泌。CRP由五个完全相同的球形单体以非共价键结合组成稳定的盘状结构，属于正五聚体家族。CRP在结构上为对称的五面体，其单体由206个氨基酸组成，分子量约为23KDa，CRP的总分子量约为118KDa。CRP参与体内的各种炎症及免疫反应，是冠状动脉粥样硬化性心脏病(冠心病)的重要标志物。大量研究表明：高水平的CRP是周围性血管病、心肌梗死、脑血管病及血管性死亡的独立风险预测因子。
在正常人血清中CRP含量极少，浓度为0.062-8.2mg/L，半衰期约为15小时。当发生细菌感染2小时后即开始升高，平均8小时增加1倍，48小时达高峰，其在病毒感染时则不会升高。CRP持续升高则提示机体存在慢性炎症或自身免疫性疾病。CRP变化不受病人的个体差异、机体状态及治疗药物的影响。另外，CRP在其他领域亦发挥着重要的作用：①在由慢性炎症引发心血管疾病的独立危险因素中CRP的作用已被证实，及时监测CRP的水平变化对心血管疾病的干预及预后起重要作用。有些学者甚至认为CRP可作为心血管危险评估的金标准，且CRP水平越高，发生心脑血管的危险性就越大；②CRP可用来评价急性胰腺炎的严重程度，当发生广泛坏死性胰腺炎时血清中CRP的水平可高达250mg/L；③如将CRP与甲胎蛋白联合应用，可用来鉴别肝脏恶性肿瘤与良性疾病，为临床治疗方案制定提供指导；另外，CRP测定对肿瘤的治疗和预后亦有积极的意义，恶性肿瘤患者CRP水平大都升高，手术切除肿瘤后CRP水平则会下降，且进行放疗、化疗和皮质激素治疗对血清CRP的影响非常小，所以测定血清CRP的水平变化有助于临床估测各个器官系统中恶性肿瘤的进展过程。④评估患者预后：CRP水平越高，提示疾病控制不佳，预后不良。
鉴于CRP在众多领域中的重要作用，制备CRP抗体并用于制备CRP定量检测试剂盒具有重要的临床应用价值。CRP通常是通过抗体浊度法或免疫比浊法测定的，而他们的检测能力在3-5mg/L以上，这个水平仅仅适用于对感染的预测，而对于冠脉及脑血管的危险预测是远远不够的。除上述技术以外，使用胶体金和荧光定量检测技术的CRP试纸卡，可满足快速检测及床旁检测的需求。其中基于胶体金定量层析技术的全程CRP检测方法既具有全自动生化仪免疫透射比浊法试剂的准确性，同时检测更加简便、快速，适合床旁诊断，易于在门诊、急诊或者ICU病房内开展。
发明内容
本发明要解决的技术问题是提供能有效的、特异性结合人C-反应蛋白的抗体。更具体地说：
本发明的第一目的在于提供两种抗人C-反应蛋白抗体。
第一种抗人C-反应蛋白抗体(P20)，
其重链可变区含有以下的互补决定区：氨基酸序列如序列SEQ ID NO：1所示的HCDR1、如序列SEQ ID NO：2所示的HCDR2和/或如序列SEQ ID NO：3所示的HCDR3；
其轻链可变区序列含有以下的互补决定区：氨基酸序列如序列SEQ ID NO：4所示的LCDR1、如序列SEQ ID NO：5所示的LCDR2和/或如序列SEQ ID NO：6所示的LCDR3。
优选的是本发明中的抗体P20的重链可变区的氨基酸序列如SEQ ID NO:7所示，轻链可变区的氨基酸序列如SEQ ID NO:8所示。
第二种抗人C-反应蛋白抗体(P02)，
其重链可变区含有以下的互补决定区：氨基酸序列如序列SEQ ID NO：9所示的HCDR1、如序列SEQ ID NO：10所示的HCDR2和/或如序列SEQ ID NO：11所示的HCDR3；
其轻链可变区序列含有以下的互补决定区：氨基酸序列如序列SEQ ID NO：12所示的LCDR1、如序列SEQ ID NO：13所示的LCDR2和/或如序列SEQ ID NO：14所示的LCDR3。
优选的是本发明中抗体P02的重链可变区的氨基酸序列如SEQ ID NO:15所示，抗体轻链可变区的氨基酸序列如SEQ ID NO:16所示。
本发明第二个目的是提供两种单链抗体，所述单链抗体P20的氨基酸序列如SEQID NO:17所示；所述单链抗体P02的氨基酸序列如SEQ ID NO:18所示。
本发明第三个目的是提供两种编码上述单链抗体的核苷酸序列，编码单链抗体P20的核苷酸序列如SEQ ID NO:19所示，和编码单链抗体P02的核苷酸序列如SEQ ID NO:20所示。
本发明第四个目的是提供一种含有上述核苷酸序列的表达载体。
本发明第五个目的是提供一种含有上述表达载体的重组宿主细胞。所属宿主细胞可以是大肠杆菌、酵母或哺乳动物细胞，优选为大肠杆菌。
本发明第六个目的是提供一种生产上述单链抗体的方法，包括：
1)在合适的条件下培养上述重组宿主菌表达抗体；
2)然后从宿主菌中纯化、收集抗体。
本发明的第七个目的在于提供上述抗人C-反应蛋白抗体在检测人C-反应蛋白含量中的应用。
本发明的第八个目的在于提供一组可进行配对并检测人C-反应蛋白的抗体对组合P20和P02；该抗体对组合的检测灵敏度高，特异性好。
本发明的第九个目的在于提供一种利用所述抗人C-反应蛋白抗体检测人C-反应蛋白的胶体金免疫层析定量检测卡，包括样品吸收垫、金标垫、反应膜和吸水垫；所述金标垫喷涂有胶体金颗粒标记的抗体P02或P20，所述反应膜上有检测带和质控带，检测带位置包被有抗体P20或P02，质控带位置包被抗His标签抗体或Protein L。所述反应膜优选硝酸纤维素膜。所述抗His标签抗体优选鼠抗His抗体。
本发明制备了多种抗体，并进行配对筛选，获得灵敏度及特异性均能满足需求的抗体组合(P20及P02)；同时其方便大量生产，可满足日后大规模临床应用的需求。对上述抗体组合进行检测体系的调试优化工作，获得操作简便，灵敏度，特异性及相关检测性能均可满足人临床样本检测的人C-反应蛋白的胶体金免疫层析定量检测卡。
附图说明
图1：PCR鉴定P20和P02抗体重链及轻链可变区琼脂糖凝胶电泳图。
其中，Lane 1为200bp DNA Ladder；Lane 2为抗体P20重链可变区DNA序列；Lane 3为抗体P20轻链可变区DNA序列；Lane 4为抗体P02重链可变区DNA序列；Lane 5为抗体P02轻链可变区DNA序列。
图2：单链抗体结构示意图。VH表示重链可变区序列，VL表示轻链可变区序列，His标签为六个组氨酸。
图3：单链抗体P20和P02在重组大肠杆菌工程菌中的诱导表达鉴定图。
其中，图3-a为单链抗体P20和P02在重组大肠杆菌工程菌中的诱导表达SDS-PAGE凝胶电泳图。泳道1为(10-230kDa)宽范围的预染的蛋白上样Marker；泳道2为未加入IPTG诱导的单链抗体P20重组大肠杆菌工程菌裂解液；泳道3为加入IPTG诱导的单链抗体P20重组大肠杆菌工程菌裂解液；泳道4为未加入IPTG诱导的单链抗体P02重组大肠杆菌工程菌裂解液；泳道5为加入IPTG诱导的单链抗体P02重组大肠杆菌工程菌裂解液。
图3-b为单链抗体P20和P02在重组大肠杆菌工程菌中的诱导表达蛋白免疫印迹图。泳道1为(10-230kDa)宽范围的预染的蛋白上样Marker；泳道2为未加入IPTG诱导的单链抗体P20重组大肠杆菌工程菌裂解液；泳道3为加入IPTG诱导的单链抗体P20重组大肠杆菌工程菌裂解液；泳道4为未加入IPTG诱导的单链抗体P02重组大肠杆菌工程菌裂解液；泳道5为加入IPTG诱导的单链抗体P02重组大肠杆菌工程菌裂解液。
图4：单链抗体P20和P02重组大肠杆菌工程菌裂解液经过HisTrap HP亲和柱纯化收集峰合并样品SDS-PAGE凝胶电泳图。泳道1为(10-230kDa)宽范围的预染的蛋白上样Marker；泳道2为单链抗体P20重组大肠杆菌工程菌裂解液经过HisTrap HP亲和柱纯化收集峰合并样品；泳道3为单链抗体P02重组大肠杆菌工程菌裂解液经过HisTrap HP亲和柱纯化收集峰合并样品。
图5：本发明胶体金免疫层析定量检测卡结构示意图。1为样品垫、2为反应膜、3为吸收垫、4为质控线(C线)、5为检测线(T线)、6为金标垫、7为PVC片材。
图6：CRP胶体金检测卡检测范围拟合曲线。
图7：CRP胶体金检测卡线性范围拟合曲线。
图8：CRP胶体金检测卡与对照产品比对拟合曲线。
具体实施方式
定义
“抗体”又称免疫球蛋白，是一类由B淋巴细胞分泌的大型Y形蛋白质，能够通过Y形的其中两个分叉顶端的互补位点(抗原结结合位)特异性结合靶抗原的免疫球蛋白分子，所述靶抗原如蛋白质、糖、多核苷酸、脂、多肽、小分子化合物等。
“单链抗体”(scFv)指的是抗体的重链可变区(VH)和轻链可变区(VL)通过15～20个氨基酸短肽(linker)连接形成的单一链融合蛋白，用于连接的linker通常富含甘氨酸和丝氨酸，以利于单链抗体的稳定性与柔韧性。连接方式可将VL的N端连接至VH的C末端，或者相反。尽管去除了恒定区并引入linker，单链抗体依然保留了抗体对抗原的特异性，且其具有分子量小、穿透力强和抗原性弱等特点。
互补决定区(complementarity-determining region，CDR)，也叫做高变区。成型于抗体单体氨基酸的末端，是靶抗原与抗体结合的最关键区域，在免疫网络理论中，每个抗体的互补决定区又被称为独特型或者基因型。
实施例1.抗人C-反应蛋白杂交瘤细胞株的制备
1.动物免疫
以提取于人血浆的C-反应蛋白(购自HyTest公司)按照一般免疫程序免疫BALB/c雌性小鼠(购自常州卡文斯实验动物有限公司)。具体免疫情况参见《抗体制备与使用实验指南》。采用间接ELISA法跟踪免疫小鼠血清滴度，选取血清效价最高的免疫小鼠，进行小鼠脾细胞和小鼠骨髓瘤细胞进行融合实验。
2.细胞融合
(1).脾脏细胞的制备
将免疫小鼠，摘眼球取血，经断颈椎处死后置于75％(v/v)的酒精中浸泡10min，于无菌操作台中取出其脾脏，置于细胞筛网中，充分研磨细胞，过筛网，用无菌1640培养基(购自Gibco公司)离心洗涤数次后，重悬细胞以制成单细胞悬液，并计数，备用。
(2).饲养细胞的制备
取8～10周龄的雌性BALB/c小鼠一只，摘眼球获取阴性血清，经断颈椎处死后置75％(v/v)酒精中浸泡10min；无菌揭开腹部皮肤，暴露腹膜，用注射器将约10mL 1640HT培养基(购自SIGMA公司)注入小鼠腹腔，轻轻按摩腹部并吹打数次。吸取含有巨噬细胞的培养基注入20％1640HAT培养基中备用；
取2～3周龄的雌性BALB/c小鼠一只，经断颈椎处死后置于75％(v/v)酒精中浸泡10min；无菌取胸腺于细胞筛网中，研磨，过筛网，获得胸腺细胞置于上述含有巨噬细胞的20％1640HAT培养基中，备用。
(3).细胞融合
选择处于对数生长期的小鼠骨髓瘤细胞株SP2/0，收集并计数。取约108个上述脾细胞与2×107个上述SP2/0细胞株加入融合管中混合，1000rpm离心10分钟后弃上清(尽量弃净)，将融合管置手掌上来回轻轻摩擦以使沉淀松散。60秒内先慢后快地加入1mL预热的PEG1450(聚乙二醇1450，购自SIGMA公司)，加入1640HT培养基30mL终止，1000rpm离心10分钟，去上清，轻轻摩擦使沉淀松散，加入步骤2所获得的20％的1640HAT培养基中。
将上述HAT培养基充分混匀后，以200μL/孔分装至96孔细胞培养板中，置37℃，5％CO2的细胞培养箱中培养。一周后用10％1640HT培养基替换20％1640HAT培养基，3天后取上清进行检测。
3.抗人C-反应蛋白特异性杂交瘤株筛选
(1).检测板的准备：用CB包被液稀释CRP(购买于HyTest公司)至1μg/mL，包被96孔ELISA酶标板，100μL/孔，2～8℃包被过夜，洗涤一次拍干；含2％牛血清白蛋白(BovineSerum Albumin，BSA，购买于盐城赛宝生物科技有限公司)的PBST缓冲液封闭(200ul/孔)，37℃封闭2小时；拍干，备用。
(2).阳性克隆的筛选：将待检细胞培养上清100μL/孔加入上述检测板中，于37℃作用30分钟后洗涤并拍干，加入100μL/孔的HRP标记的羊抗鼠IgG，于37℃作用30分钟后洗涤并拍干，加入100μL/孔的TMB显色液，于37℃避光显色15分钟，每孔加入50μL的2M H2SO4终止反应，并于OD450处读取数值。阳性孔确定原则：OD450值/阴性对照值≥2.1。选取阳性克隆株进行细胞克隆化筛选。经过三至四轮的克隆化筛选后，单克隆细胞株阳性率100％即确定为稳定细胞株，对细胞株进行定株。杂交瘤细胞株M20及M02均具有较高的效价，遂后续进一步对上述杂交瘤细胞株进行抗体可变区序列测序分析。
实施例2.杂交瘤细胞株抗体可变区序列的测定
对上述杂交瘤细胞株M20及M02抗体可变区序列进行测定。
a.RNA的提取：参照细胞总RNA抽提试剂盒(购自Roche公司)说明书对上述杂交瘤细胞株M20及M02进行总RNA提取并立即进行反转录；
b.RNA反转录成为DNA：参照Thermo Scientific Reverted First strand cDNASynthesis Kit(购自Thermo公司)对上一步骤中所提取的总RNA进行反转录，制得cDNA，冻存于-20℃备用；
c.可变区序列的PCR扩增及回收：以上一步骤中所得cDNA为模板，以鼠IgG亚型单克隆抗体可变区序列通用引物为引物，对重链及轻链的可变区序列进行PCR扩增，将PCR产物经DNA胶回收试剂盒(购自TIANGEN公司)进行回收，见附图1；
d.可变区序列的克隆和序列测定：按照克隆载体pMD18-T kit(购自Takara公司)说明书，将重链和轻链可变区基因分别与pMD18-T载体进行连接，转化大肠杆菌DH5α，挑取阳性克隆，交由南京金斯瑞生物科技有限公司进行测序。
测序得到杂交瘤细胞株M20的抗体(记为P20)重链可变区氨基酸序列如SEQ IDNO:7所示、轻链可变区氨基酸序列如SEQ ID NO：8所示。Vbase2数据库分析上述序列，其重链可变区的各互补决定区的氨基酸序列分别是：如序列SEQ ID NO：1所示的HCDR1、如序列SEQ ID NO：2所示的HCDR2和/或如序列SEQ ID NO：3所示的HCDR3；其轻链可变区的各互补决定区的氨基酸序列是：如序列SEQ ID NO：4所示的LCDR1、如序列SEQ ID NO：5所示的LCDR2和/或如序列SEQ ID NO：6所示的LCDR3。
测序得到杂交瘤细胞株M02的抗体(记为P02)重链可变区氨基酸序列如SEQ IDNO:15所示、轻链可变区氨基酸序列如SEQ ID NO：16所示。Vbase2数据库分析上述序列，其重链可变区的各互补决定区的氨基酸序列分别是：如序列SEQ ID NO：9所示的HCDR1、如序列SEQ ID NO：10所示的HCDR2和/或如序列SEQ ID NO：11所示的HCDR3；其轻链可变区的各互补决定区的氨基酸序列是：如序列SEQ ID NO：12所示的LCDR1、如序列SEQ ID NO：13所示的LCDR2和/或如序列SEQ ID NO：14所示的LCDR3。
实施例3.单链抗体的重组表达及纯化
根据实施例2中测序结果，分别将M20及M02抗体重链及轻链可变区之间加入连接肽(GGGGS)3，并将其全基因进行合成，进行单链抗体的表达载体构建和重组表达。所表达得到的抗体分别命名为抗体P20及抗体P02，其结构组成如附图2所示。上述单链抗体的重组表达具体如下：
a)单链抗体P20和P02表达质粒构建
单链抗体P20的核苷酸序列如SEQ ID NO:19所示、氨基酸序列如SEQ ID NO:17，单链抗体P02的核苷酸序列如SEQ ID NO:20所示、氨基酸序列如SEQ ID NO:18所示。将单链抗体P20和P02全基因构建到pUC57质粒(由南京金斯瑞生物科技有限公司提供)中，得到一种长期保存质粒，质粒分别记为pUC57-P20-scFv-(HIS)6和pUC57-P02-scFv-(HIS)6。进行PCR扩增，其中扩增单链抗体P20的上下游引物为P1和P2，扩增单链抗体P02的上下游引物为P3和P4
上游引物P1：CTAGCCATGGAAGTTATGTTGGTTGAATC(SEQ ID NO:21)；
下游引物P2：CCGCTCGAGTTACTAGTGATGGTGATGG(SEQ ID NO:22)。
常规PCR程序后，琼脂糖凝胶电泳分析，显示两种产物大小与预期大小一致。将PCR获得基因产物回收纯化后，采用NcoI(#R0193S,购自New England BioLabs公司)和XhoI(#R0146S,购自New England BioLabs公司)双酶切，用T4连接酶连接到pET28a(69864，购自Merck公司)质粒中，转化到DH5a感受态细胞(CB101，购自北京天根生化科技有限公司)中，在含有卡那霉素(0408，购自Amresco公司)的LB平板中37℃培养过夜。第二天筛选阳性克隆菌测序，比对，与预期序列完全一致，即得到单链抗体P20的表达质粒，记为pET28a-P20-scFv-(HIS)6和pET28a-P02-scFv-(HIS)6。
b)单链抗体P20和P02重组大肠杆菌工程菌株的构建、筛选及表达
具体步骤如下：
将实施例3步骤a中测序比对正确的pET28a-P20-scFv-(HIS)6和pET28a-P02-scFv-(HIS)6质粒转化到大肠杆菌BL21(DE3)感受态菌株(CB105，购自北京天根生化科技有限公司)中，37℃卡那霉素平板中过夜培养。第二天分别挑阳性菌落，接入含有50μg/mL卡那霉素的LB培养液，37℃培养过夜。取50μL过夜培养物接入5mL含50μg/mL卡那霉素的LB诱导培养液，37℃振荡培养。待OD600＝1.0时，用1mmol/L的IPTG(购自Amresco公司)进行诱导表达。同时以未加入IPTG的大肠杆菌培养液做阴性对照。4h后12000rpm，3min收集菌液，用预冷的PBS清洗沉淀，加入5×SDS凝胶加样缓冲液，100℃加热10min，室温12000rpm，1min高速离心，取上清。未加入IPTG的大肠杆菌培养液也按此步骤处理。各取5μL所得的未加入IPTG和加入IPTG诱导的样品，12％SDS-PAGE凝胶电泳和蛋白免疫印迹分析表达效果，Westernblot中一抗为抗HIS-Tag抗体(His-Tag(2A8)Mouse mAb，M20001，购于上海艾比玛特生物医药有限公司)见图3-a和图3-b。
c)单链抗体P20和P02纯化
采用组氨酸标签亲和柱纯化单链抗体P20和P02，预装柱子选择为HisTrap HP，具体步骤如下：
取单链抗体P20和P02重组大肠杆菌工程菌株甘油管按照1％接种量接种于含有50μg/ml卡那霉素的LB液体培养基中，37℃，220rpm培养至OD600≈1.0时加入终浓度为1mMIPTG进行诱导表达，诱导表达4h后离心收集菌体。用预冷的PBS重悬，于4℃以12000rpm，离心15min；重复一次。吸去上清，称菌体沉淀重量，每克(菌体湿重)加入5mL结合缓冲液：300mM NaCl，20mM NaH2PO4，10mM Imidazole，pH＝7.5。充分混匀后，每克(菌体湿重)菌体加入5μL 100mmol/L PMSF，5μL 100mg/mL溶菌酶，冰上孵育20min。样品置于冰上，用探针型超声波仪破碎菌体，超声120次，每次5s间隔5s，循环三次，每次循环至冷却样品之间等待2min，等待样品冷却。4℃，12000rpm，离心15min，过0.45μm滤膜。
HisTrap HP亲和柱纯化：运用全自动智能蛋白纯化系统(AKTA avant150，购自GEhealthcare公司)对上述预处理获得的单链抗体P20和P02细胞裂解液分别进行亲和纯化，柱子为HisTrap HP(17-5248-02,购自GE healthcare公司)。结合缓冲液为300mM NaCl，20mM NaH2PO4，10mM Imidazole，pH7.5，洗脱缓冲液为300mM NaCl，20mM NaH2PO4，500mMImidazole，pH7.5。洗脱时进行线性洗脱，并收集各个洗脱峰。
通过SDS-PAGE电泳鉴定纯度，合并纯度较高符合要求的单链抗体P20和P02收集管，更换缓冲液为PBS溶液并超滤浓缩(1mg/ml)，过滤除菌于-20℃保存备用。
本领域技术人员知晓，重组蛋白可以不带标签，也可带其他标签，也可加入其他形式的连接肽。无论是否带标签或者带不同形式的标签都可采用protein L亲和填料纯化。
实施例4.单链抗体的性能评价
1.单链抗体P20及P02的Western blot鉴定
a.聚丙烯酰胺凝胶电泳：分别配置12％非变性聚丙烯酰胺凝胶和SDS-聚丙烯酰胺凝胶；分别上样标准蛋白质及天然CRP蛋白(购买于HyTest公司)，恒压下电泳1小时；
b.转膜：恒流(35mA/膜)条件下转膜1小时，将聚丙烯酰胺凝胶上的蛋白质分别转移至硝酸纤维素膜上。考马斯亮蓝G250对完成转膜的聚丙烯酰胺凝胶进行染色，观察蛋白的残留情况；
c.封闭：含5％脱脂奶的TBST缓冲液封闭(封闭液)，4℃过夜；封闭后洗涤液(TBST，详见TaKaRa公司TBST buffer)洗涤三次，每次5min；
d.抗原抗体反应：封闭液稀释(按1:400体积比)辣根过氧化物酶标记P20(P20-HRP，1mg/mL，本公司采用经典过碘酸钠法标记，下同)及辣根过氧化物酶标记P02(P02-HRP，1mg/mL，本公司采用经典过碘酸钠法标记，下同)，分别加入上述硝酸纤维素膜中，室温反应1h；TBST洗涤5次，每次5min；
e.显色及拍照：吸干硝酸纤维素膜上残留液体，每张硝酸纤维素膜分别加入2mL稳定型过氧化物酶溶液(1mL)与鲁米诺/增强剂溶液(1mL)的混合液(购买于Thermo公司)，均匀润湿硝酸纤维素膜的表面，室温避光反应一分钟后于凝胶成像系统(购买于GE公司)拍照，留取结果。
实验结果表明，本发明两种抗体均可与变性及非变性血液提取的天然CRP反应，证明了本发明两种抗体与天然CRP的结合力。
2.单链抗体P20及P02在胶体金检测平台的评价
将实施例3中纯化的抗体进行配对组合，分别作为包被抗体或标记抗体进行配对检测C-反应蛋白(CRP)，检测步骤如下：
1)用抗体包被液将P20或单链抗体P02稀释至1mg/ml，划线于硝酸纤维素膜上；
2)用0.01M PB缓冲液将胶体金标记的P20或P02稀释5倍后铺于结合垫上；
3)按附图5所示贴膜，切条，装卡(具体制备参见实施例5)
4)用样本稀释液稀释CRP标准品(购买于HyTest公司)，至浓度为50ug/mL，1ug/mL，将这两个浓度标准品和零浓度标准品(即样本稀释液)分别添加50ul到胶体金检测卡中(P20包被-P02标记或P02包被-P20标记)，10min后将检测卡放置在读数仪上进行读数。结果如下表所示：
由上述结果可知，可见P20作为包被抗体，P02作为标记抗体组成的双抗体夹心法检测系统可应用于胶体金检测平台，进行天然CRP的检测。
实施例5抗人C-反应蛋白的胶体金免疫检测卡的制备
1、溶液配制
1)0.01M PB缓冲液制备：称取Na2HPO4·12H2O 3.22g，NaH2PO4·2H2O 0.15g，加纯化水1000mL，转子搅拌至溶解，用ph计测定其pH7.4±0.1，0.45um滤膜过滤。
2)封闭液制备：称取牛血清白蛋白5g，加0.01M PB溶液(PH7.4±0.1)50mL，转子搅拌至溶解。
3)抗体包被液制备：取0.2mL异丙醇，加入9.8mL 0.01M PB溶液(PH7.4±0.1)，转子搅拌5-10min。
4)金标抗体复溶液制备：称取1g牛血清白蛋白，5g海藻糖，加入100mL 0.01M PB溶液(pH7.4±0.1)，转子搅拌至溶解后，加入25ul Tween-20，继续用转子搅拌5-10min。
5)样本稀释液制备：称取12.5g牛血清白蛋白加入500mL 0.01M PB溶液(PH7.4±0.1)，转子搅拌至溶解，加入0.5ml Proclin300，继续用转子搅拌5-10min。
2、人C-反应蛋白胶体金检测卡制备
1)胶体金的标记
抗体P02的胶体金标记(以1ml胶体金溶液标记为例)：用K2CO3调节胶体金pH值(每1ml胶体金中加入5ul 0.2M K2CO3)，搅拌5-10min，向胶体金溶液中缓慢加入抗体P02(每1ml胶体金中加入25ug抗体P02)，低速搅拌30min；加入封闭液BSA至其终浓度为10％(质量百分比)，搅拌20min；静置30min后12000rpm离心30min；去上清用100uL金标抗体复溶液复溶沉淀即得胶体金标记的P02抗体。
2)金标垫及反应膜制备
将P02金标抗体稀释5倍后，喷涂在金标垫6上，干燥备用；
将抗体P20用抗体稀释液稀释至1.5mg/mL后，包被于反应膜2(硝酸纤维素膜)的T线5位置；将抗His标签抗体用抗体稀释液稀释至0.2mg/mL后，包被于反应膜2(硝酸纤维素膜)的C线4位置，反应膜干燥备用。
3)贴膜、切膜、组装
将样品垫1、金标垫6、包被有抗体的硝酸纤维素膜2、吸水垫3从左至右依次设置(如图5所示)，且两两之间应少许接触，所述包被有抗体的硝酸纤维素膜的T线5在左、C线4在右，并根据外壳大小进行切割，装入外壳，完成检测卡制备。
4)试剂盒组装
将组装好的检测卡，干燥剂装入铝箔袋中，热封机封口，贴标签；
将样本稀释液按1mL/管进行分装，并按照试剂盒规格装入自封袋，贴标签；
按照成品规格，将一定份数的内包，1个含样本稀释液的自封袋，1份说明书，1个合格标签装入包装盒内，并在包装盒外贴好标签。
3、人C-反应蛋白胶体金检测卡使用方法
1)打开外包装，从密封铝箔袋中取出检测卡，置于平坦台面上。
2)吸取2μl血清/血浆样本，加入样本稀释液中，充分混合。
3)吸取50ul经处理后的样本加入检测卡的加样孔中，室温下静置10min。
4)将检测卡放入免疫层析定量分析仪中，按“快速检测”键开始检测，仪器将自动对检测卡进行扫描。
5)从免疫层析定量分析仪的显示屏幕上读取/打印检测结果。
4、人C-反应蛋白胶体金检测卡检测效果评估
1)精密性：将P20(包被)-P02(标记)检测卡按检测卡使用方法检测1、10、50ug/ml的CRP参考品各10次重复测定，剔除离群值后计算检测卡精密度。实验结果显示三个浓度检测结果变异系数CV<15％。
2)检测范围：将P20(包被)-P02(标记)检测卡检测不同浓度的CRP重组蛋白0.5、1.56、3.125、6.25、12.5、25、50、100ug/mL，拟合曲线及检测范围为0.5-100ug/ml(如附图6)。
3)线性范围：将高值样本与样本稀释液按照一定比例配制成5个系列浓度样本0.5、25、50、75、100ug/mL，用P20(包被)-P02(标记)检测卡检测，每个样本检测3次，将结果与理论浓度进行回归统计，判断在该浓度范围内是否成线性。线性范围为0.5-100ug/ml(如附图7)。灵敏度0.5ug/mL。
4)准确度：将P20(包被)-P02(标记)检测卡按检测卡使用方法检测1、10、50ug/ml的CRP参考品各3次重复，计算平均值和理论值的相对偏差，实验结果显示三个浓度检测结果相对偏差B<15％。
5、准确度-方法学比对：
选择同类产品中目前市场上获得良好信誉的广州万孚生物技术股份有限公司全程C-反应蛋白(hsCRP+常规CRP)定量检测试剂(免疫层析法)作对照产品比对验证。选择20份临床病人标本，按1到20的顺序编号，用对照产品和待评价的P20(包被)-P02(标记)的胶体金检测卡同时进行实验，按照1，2，3......18，19，20，20，19，18......3，2，1的样本顺序进行测定。对照和待评价产品的检测结果相关系数R2＝0.980，说明两种方法检测结果有较好的相关性(如附图8)。
6、配方筛选
除上述最优制备例1外，申请人还尝试多种制备方案，例如下面几组检测卡制备及应用结果如下表：
SEQUENCE LISTING
<110> 江苏众红生物工程创药研究院有限公司
<120> 一种抗人CRP抗体及其应用
<130> 一种抗人CRP抗体及其应用
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Ser Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys
195 200 205
Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln
210 215 220
Asn Thr His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile
225 230 235 240
Lys Arg
<210> 18
<211> 242
<212> PRT
<213> 人工序列
<400> 18
Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ala Met Ser Trp Val Arg Gln Thr Pro Glu Arg Arg Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Thr Ser Gly Ser Asp Arg Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Trp Arg Phe Thr Ile Ser Arg Asp Ser Ala Lys Asn Ile Leu Tyr
65 70 75 80
Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Arg Arg Gly Trp Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val
100 105 110
Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
115 120 125
Ser Asp Val Leu Met Thr Gln Arg Pro Leu Ser Leu Pro Val Ser His
130 135 140
Gly Asp Gln Ala Ser Thr Ser Cys Arg Ser Ser Gln Ser Ile Val His
145 150 155 160
Ser Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln
165 170 175
Ser Pro Lys Leu Leu Ile Tyr Lys Val Phe Asn Arg Phe Ser Gly Val
180 185 190
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys
195 200 205
Ile Ser Arg Val Glu Ala Glu Asp Gln Gly Val Tyr Tyr Cys Phe Gln
210 215 220
Gly Ser Arg Asp Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile
225 230 235 240
Lys Arg
<210> 19
<211> 726
<212> DNA
<213> 人工序列
<400> 19
gaagttatgt tggttgaatc tggtggtggt ttggttaagc caggtggttc tttgaaattg 60
tcttgtgctg cttctggttt tactttctct tcttacgcta tgtcttgggt tagacaaact 120
ccagagagaa gattggaatg ggttgctact atttctactt ctggttctga tagatactat 180
cctgattctg ttaagtggag attcactatt tccagagatt ctgctaaaaa catcttgtac 240
ttgcaaatgt cttctttgag atctgaggat actgctatgt actattgtgc tagaagaggt 300
tgggattatt ggggtcaagg tacttctgtt actgtttctg ctggtggtgg tggttccgga 360
ggtggtggtt ctggtggtgg tggttctgat gttgttatga cccaaactcc attgtctttg 420
cctgtttctt tgggagatca agcttctatt tcttgtagat cttctcaatc tttggttcat 480
tctaacggta atacttactt gcactggtat ttgcaaaagt ctggtcaatc tcctaagttg 540
ttgttgtaca aagtttctaa cagattttct ggtgtttctg atagattctc tggttctggt 600
tctggtactg atttcacttt gaagatttcc agagttgagg ctgaagattt gggtgtttac 660
ttctgttctc aaaacactca tgttccacct actttcggtg gtggtactaa gttggaaatt 720
aaaaga 726
<210> 20
<211> 726
<212> DNA
<213> 人工序列
<400> 20
gaagttatgt tggttgaatc tggtggtggt ttggttaagc caggtggttc tttgaaattg 60
tcttgtgctg cttctggttt tactttctct tcttacgcta tgtcttgggt tagacaaact 120
ccagagagaa gattggaatg ggttgctact atttctactt ctggttctga tagatactat 180
cctgattctg ttaagtggag attcactatt tccagagatt ctgctaaaaa catcttgtac 240
ttgcaaatgt cttctttgag atctgaggat actgctatgt actattgtgc tagaagaggt 300
tgggattatt ggggtcaagg tacttctgtt actgtttctt ctggtggtgg tggatccgga 360
ggtggtggtt ctggtggtgg tggttctgat gttttgatga cccaaagacc attgtctttg 420
cctgtttctc atggagatca agcttctact tcttgtagat cttctcaatc tattgttcac 480
tctaacggta atacttactt ggagtggtat ttgcaaaagc caggtcaatc tcctaagttg 540
ttgatctaca aggtttttaa tagattctct ggtgttcctg atagattttc tggttctggt 600
tctggtactg atttcacttt gaagatttcc agagttgagg ctgaagatca aggtgtttac 660
tattgttttc aaggttccag agatccacct actttcggtg gtggtactaa gttggaaatt 720
aaaaga 726
<210> 21
<211> 29
<212> DNA
<213> 人工引物
<400> 21
ctagccatgg aagttatgtt ggttgaatc 29
<210> 22
<211> 28
<212> DNA
<213> 人工引物
<400> 22
ccgctcgagt tactagtgat ggtgatgg 28
Claims (8)
1.抗人C-反应蛋白抗体，其重链可变区含有以下的互补决定区：氨基酸序列如序列SEQID NO：1所示的HCDR1、如序列SEQ ID NO：2所示的HCDR2和/或如序列SEQ ID NO：3所示的HCDR3；
其轻链可变区序列含有以下的互补决定区：氨基酸序列如序列SEQ ID NO：4所示的LCDR1、如序列SEQ ID NO：5所示的LCDR2和/或如序列SEQ ID NO：6所示的LCDR3。
2.权利要求1所述的抗人C-反应蛋白抗体，其特征在于重链可变区的氨基酸序列如SEQID NO:7所示，轻链可变区的氨基酸序列如SEQ ID NO:8所示。
3.抗人C-反应蛋白单链抗体，所述单链抗体的氨基酸序列如SEQ ID NO:17所示。
4.编码权利要求书3所述单链抗体的核苷酸序列，如SEQ ID NO:19所示。
5.含有权利要求4所述核苷酸序列的表达载体。
6.含有权利要求5所述表达载体的重组宿主细胞。
7.生产权利要求3所述单链抗体的方法，包括：
1)在合适的条件下培养上述重组宿主菌表达抗体；
2)然后从宿主菌中纯化、收集抗体。
8.权利要求1至3任一项所述抗人C-反应蛋白抗体在检测人C-反应蛋白含量中的应用。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112574304A (zh) * | 2019-09-30 | 2021-03-30 | 东莞市朋志生物科技有限公司 | 一种抗c反应蛋白的抗体 |
| CN114773462A (zh) * | 2022-04-12 | 2022-07-22 | 内蒙古农业大学 | 用于检测牛crp蛋白的重组单链抗体及其应用 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202794189U (zh) * | 2012-08-07 | 2013-03-13 | 天津中新科炬生物制药有限公司 | C反应蛋白快速全程定量免疫层析检测试剂盒 |
| CN105158476A (zh) * | 2015-06-03 | 2015-12-16 | 南京闻智生物科技有限公司 | 一种全量程c反应蛋白胶乳增强免疫比浊检测试剂盒 |
| CN105949309A (zh) * | 2016-04-12 | 2016-09-21 | 江苏众红生物工程创药研究院有限公司 | 抗人c-反应蛋白抗体及其应用 |
-
2017
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202794189U (zh) * | 2012-08-07 | 2013-03-13 | 天津中新科炬生物制药有限公司 | C反应蛋白快速全程定量免疫层析检测试剂盒 |
| CN105158476A (zh) * | 2015-06-03 | 2015-12-16 | 南京闻智生物科技有限公司 | 一种全量程c反应蛋白胶乳增强免疫比浊检测试剂盒 |
| CN105949309A (zh) * | 2016-04-12 | 2016-09-21 | 江苏众红生物工程创药研究院有限公司 | 抗人c-反应蛋白抗体及其应用 |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112574304A (zh) * | 2019-09-30 | 2021-03-30 | 东莞市朋志生物科技有限公司 | 一种抗c反应蛋白的抗体 |
| CN114773462A (zh) * | 2022-04-12 | 2022-07-22 | 内蒙古农业大学 | 用于检测牛crp蛋白的重组单链抗体及其应用 |
| CN114773462B (zh) * | 2022-04-12 | 2023-07-07 | 内蒙古农业大学 | 用于检测牛crp蛋白的重组单链抗体及其应用 |
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