CN113980125B - 一种抗sftsv的中和性单克隆抗体及其应用 - Google Patents
一种抗sftsv的中和性单克隆抗体及其应用 Download PDFInfo
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- CN113980125B CN113980125B CN202111212461.1A CN202111212461A CN113980125B CN 113980125 B CN113980125 B CN 113980125B CN 202111212461 A CN202111212461 A CN 202111212461A CN 113980125 B CN113980125 B CN 113980125B
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Abstract
本发明涉及一种抗SFTSV的中和性单克隆抗体，其重链可变区包含三个CDR区，所述CDR区的氨基酸序列分别如SEQ ID NO:1、SEQ ID NO:2和SEQ ID NO:3所示，轻链可变区包含三个CDR区，所述CDR区的氨基酸序列分别如SEQ ID NO:4、SEQ ID NO:5和SEQ ID NO:6所示。本发明还提供了包含上述单克隆抗体的药物或疫苗。本发明的单克隆抗体40C10具有中和抗体活性，能够特异性识别SFTSV‑Gn蛋白的空间表位。该抗体能够中和SFTSV的四种基因型的毒株和SFTSV关联病毒GTV，可以作为SFTSV抗病毒药物以及抗体用于SFTSV和GTV的防治。
Description
技术领域
本发明涉及抗体药物技术领域，具体涉及一种抗SFTSV的中和性单克隆抗体及其应用。
背景技术
发热伴血小板减少综合征病毒(Severe fever with thrombocytopeniasyndrome，SFTSV)是新发出血热病毒，长角血蜱是主要的宿主蜱虫。该病毒在中国中东部湖北、河南、浙江、山东等地区以及日本和韩国流行。主要的感染人群为中老年人，感染后会出现高热、厌食、肌肉痛、寒战、淋巴结肿大、白细胞降低、血小板减少等症状，严重者导致多器官功能衰竭，死亡率13-30％，主要是通过蜱虫叮咬的方式传播，感染人群多为农民，也有文献报道出现过人传人的事件。
古尔图病毒(Guertu Virus，GTV)是从我国新疆的蜱虫样本中分离出来的一种病毒，目前尚未发现临床病例的报道，但血清学相关研究显示该病毒感染人的可能性。SFTSV和GTV都是以蜱虫为媒介，亲缘关系接近，均属于布尼亚病毒目，白纤病毒科，班大病毒属。目前针对该病毒引起的疾病没有有效的疫苗和治疗药物，因此开展该类病毒的中和抗体研究是十分重要的。
关于SFTSV毒株的单克隆抗体，相关研究利用SFTSV感染患者的血清抗体库平台，筛选并加工获得了针对SFTSV毒株的人源化的单克隆抗体。相关研究通过制备Gn蛋白、免疫双峰骆驼、利用噬菌体库、纳米单抗的平台技术等，筛选到特异性结合Gn的纳米抗体，鉴定了其CDR序列，并构建了人源化抗体。但是对于SFTSV引起的疾病目前并没有有效的疫苗和治疗药物。
但是目前尚无关于GTV的中和抗体研究报道，因此找到能够中和SFTSV和GTV病毒的单克隆抗体是十分重要的，对该类病毒导致的疾病防治工作具有重要意义。
发明内容
本发明为解决上述技术问题提供了一种抗SFTSV的中和性单克隆抗体，能够中和SFTV和GTV。
本发明解决上述技术问题的技术方案如下：一种抗SFTSV的中和性单克隆抗体，所述单克隆抗体的重链可变区包含三个CDR区，所述CDR区的氨基酸序列分别如SEQ ID NO:1、SEQ ID NO:2和SEQ ID NO:3所示，所述单克隆抗体的轻链可变区包含三个CDR区，所述CDR区的氨基酸序列分别如SEQ ID NO:4、SEQ ID NO:5和SEQ ID NO:6所示。
在上述技术方案的基础上，本发明还可以做如下改进。
进一步，所述重链可变区具有如SEQ ID NO:7所示的氨基酸序列，所述轻链可变区具有如SEQ ID NO:8所示的氨基酸序列。
进一步，所述单克隆抗体的重链可变区由SEQ ID NO:9所示的核苷酸序列编码，所述单克隆抗体的轻链可变区由SEQ ID NO:10所示的核酸序列编码。
进一步，所述重链的恒定区为IgG类型，所述轻链的恒定区为κ链。
本发明还提供了上述单克隆抗体在制备预防或治疗SFTSV和/或GTV感染药物中的应用。
本发明还提供了一种药物或疫苗，包括上述单克隆抗体。
进一步，所述药物或疫苗还包括药学上可接受的载体、赋形剂、稀释剂和佐剂中的一种或多种。
本发明的有益效果：本发明的单克隆抗体40C10具有中和抗体活性，能够特异性识别SFTSV-Gn蛋白的空间表位。该抗体能够中和SFTSV的四种基因型的毒株(HBGS13-C2、HBMC5-C3、WCH-C4、NB24-J)和GTV病毒。并且能够在三种毒株(HBGS13-C2、HBMC5-C3、WCH-C4)感染A129小鼠(I型干扰素α/β受体基因(IFNAR1)缺陷)模型上保护小鼠，具有广谱的中和效果。可以作为SFTSV抗病毒药物以及抗体用于SFTSV和GTV的防治。
附图说明
图1为本发明实施例1单克隆抗体40C10的蛋白免疫印迹分析结果，其中SFTSV-Vero为试验组，Vero为对照组；
图2为本发明实施例1的单克隆抗体40C10的间接免疫荧光结果，其中图2a为阳性对照组免疫荧光照片，图2b为试验组免疫荧光照片，图2c为阴性对照组一免疫荧光照片，图2d为阴性对照组二免疫荧光照片；
图3为本发明实施例1的单克隆抗体40C10的中和效价测定结果，其中图3a为试验组荧光照片，图3b为对照组荧光照片；
图4为本发明实施例2单克隆抗体40C10的蛋白免疫印迹分析结果，其中图4a使用Gn的兔多抗作为一抗的蛋白免疫印迹结果，图4b是使用40C10作为一抗的蛋白免疫印迹结果；
图5为本发明实施例2的单克隆抗体40C10的间接免疫荧光结果，其中图5a、5b和5c分别为试验组、阳性对照组和阴性对照组的结果；
图6为本发明实施例2的单克隆抗体的小鼠体内保护性结果，图6a为40C10保护C2(HBGS13)毒株攻毒A129小鼠的体重变化，图6b为40C10保护C2(HBGS13)毒株攻毒A129小鼠的生存率；图6c为40C10保护C3(HBMC5)毒株攻毒A129小鼠的体重变化，图6d为40C10保护C3(HBMC5)毒株攻毒A129小鼠的生存率；图6e为40C10保护C4(WCH)毒株攻毒A129小鼠的体重变化，图6f为40C10保护C4(WCH)毒株攻毒A129小鼠的生存率。
具体实施方式
以下结合附图及具体实施例对本发明的原理和特征进行描述，所举实例只用于解释本发明，并非用于限定本发明的范围。
实施例1杂交瘤细胞制备以及单克隆抗体筛选
1、动物免疫
将灭活的病毒SFTSV(HBMC5)用等体积的弗氏(不)完全佐剂乳化后，通过腹腔及皮下多点注射的方式对6-8周龄的SPF级雌性BALB/c小鼠进行两次免疫，每次免疫间隔两周，免疫体积为100-200μL/只。
2、制备杂交瘤细胞
取加强免疫后BALB/c小鼠，眼眶放血处死，将致死的小鼠于75％的酒精中浸泡5min，然后在超净台中使用无菌的器械解剖小鼠，取小鼠脾脏，将无菌的脾脏放在无菌的匀浆器中，加入5mL RPMI-1640培养基研磨，再补加10mL培养基充分混匀，静止10min后取研磨上清，1000rpm/min离心10min后弃上清，沉淀加入10mL培养基重悬细胞。
取免疫脾细胞悬液(1X108cells)加入骨髓瘤细胞(2X107cells),混匀后，1600rpm/min离心10min，弃上清。将装有细胞混合液的离心管放37℃水浴环境下，缓慢加入1mL预热的50％PGE，边加边混匀。然后5分钟内缓慢加入预热的RPMI-1640培养基10mL。加完以后，37℃放置10min,1000rpm/min离心8min,弃去上清。然后将细胞用50mL HAT培养基重悬，将融合后杂交瘤细胞铺在96孔板中每孔100μL，稀释后使96孔板中每个孔为单细胞状态，置于37℃，5％CO2细胞培养箱中培养7-10d，取融合细胞上清，进行单克隆抗体筛选。
3、单克隆抗体筛选
3.1单克隆抗体的蛋白免疫印迹分析(WB)
SFTSV感染于Vero细胞中扩增，经蔗糖密度梯度离心法纯化回收。将病毒蛋白经15％SDS-PAGE凝胶电泳分离，转移到0.2μm硝化纤维素膜上。用TBS-T配制的5％(w/v)脱脂奶粉37℃封闭2h，将融合细胞的上清作为一抗，在37℃孵育2h或4℃过夜。TBST洗四遍，分别加入1:5000比例稀释的HRP标记山羊抗鼠和山羊抗兔IgG或HRP标记ProteinA/G(Trans-GenBiotech，北京)，在37℃孵育1h。进行免疫印迹显示，在GE Image Quant LAS4000仪器(GEHealthcare，英国)上成像。对照组不用SFTSV感染Vero细胞，其他步骤同上。
3.2单克隆抗体的间接免疫荧光验证
将Vero细胞以1×104/孔铺至96孔细胞板，每孔加入100μL 2％胎牛血清的DMEM。待细胞密度长至70-80％时，弃掉上清，加入含SFTSV的细胞培养上清，补加至100μL含有100U/mL青霉素和100μg/mL链霉素的2％胎牛血清的DMEM。培养3d后，将细胞用无菌的PBS洗一遍，每孔加300μL4％的多聚甲醛进行固定15min。随后进行细胞透化，每孔加100μL 0.4％的TritonX-100孵育10min。每孔加100μL 5％BSA封闭，37℃孵育2h或4℃过夜。弃掉封闭液，融合细胞的培养基上清作为一抗，用1％的BSA将兔抗SFTSV-NP(稀释1:1000)作为阳性对照，37℃孵育2h或4℃过夜。分别用FITC标记的羊抗鼠和羊抗兔荧光二抗按照1:1000的比例用1％的BSA进行稀释并在每孔加入50μL，37℃孵育1h。每孔加入50μL Hoechst 33342染核液，室温放5min，荧光显微镜下观察。
另外设置三组实验作为对照，其中阳性对照组采用SFTSV的兔多抗作为一抗，其他设置同试验组；阴性对照组一健康Vero细胞采用SFTSV的兔多抗作为一抗，其他设置同试验组；阴性对照组二健康Vero细胞采用40C10抗体作为一抗，其他设置同试验组。
3.3单克隆抗体的中和效价的测定
将Vero细胞以1×104每孔铺至96孔细胞板。将单克隆抗体进行梯度稀释，取50μL单克隆抗体和50μL含100个TCID50病毒混合在一起，37℃孵育1.5h。将抗体和病毒混合液加入至96孔细胞板，37℃培养3天后进行免疫荧光检测。对照组不加单克隆抗体40C10，换为等体积的PBS，其他设置同试验组。
筛选得到单克隆抗体40C10，其蛋白免疫印迹分析结果如图1所示，阳性对照组的SFTSV的兔多抗可以识别SFTSV的Gn蛋白，对照组及实验组中均未发现有蛋白与单克隆抗体40C10结合，证明筛选得到的单克隆抗体40C10不能识别SFTSV蛋白的线性表位。单克隆抗体40C10的间接免疫荧光结果如图2所示，说明单克隆抗体40C10具有识别SFTSV病毒蛋白的活性。中和效价测定的结果如图3所示，证明单克隆抗体40C10可以中和SFTSV病毒。
实施例2、单克隆抗体40C10的鉴定
1.抗体可变区序列测定
收集培养的细胞，加入QIXzol试剂1mL，反复吹打裂解细胞后按QIXzol试剂说明书提取总RNA。使用紫外分光光度计检测所提取的RNA浓度和纯度。取3μg RNA，加入Oligo(dT)(500μg/mL)、10mM dNTP Mix、5×First-Strand Buffer、0.1M DTT、RNaseOUTTMRecombinantRibonuclease Inhibitor和M-MLV RT进行cDNA的合成。取2μL cDNA作为模板，使用高保真酶(2×Max MasterMix)进行PCR扩增获得目的基因片段，引物序列见下表1。PCR反应条件：95℃预变性3min后，扩增30个循环，每个循环条件为：94℃变性15s，55℃退火15s，72℃延伸30s，最后72℃彻底延伸5min。取30-50μLPCR产物进行1％琼脂糖电泳。
按Gel Extraction Kit说明书纯化300bp左右大小的PCR产物。以pTOPO-Blunt作为克隆载体，按照Zero BackgroundpTOPO-Blunt Cloning Kit说明书分别将轻重链可变区域目的基因与pTOPO-Blunt连接。取2μL重组子用来转化处于感受态的DH10B受体菌,然后涂布于含100μg/mL氨苄青霉素Ampicillin的LB平板上，37℃过夜。分别用新鲜灭菌枪头从平板上挑取轻重链单克隆菌各5-10个落于1mL LB培养基(含氨苄50μg/mL)中，37℃300rpm培养8-12h后送菌液用通用引物M13F和M13R测序。
表1
测序得到编码单克隆抗体40C10的重链可变区核苷酸序列SEQ ID NO:9和轻链可变区的核苷酸序列SEQ I NO:10。SEQ ID NO:9和SEQ I NO:10的序列如下所示：
SEQ IDNO:9：
gaggttcagctgcagcagtctggggctgaactggtgaagcctggggcttcagtgaagttgtcctgcaaggcttctggctacaccttcaccagccactatatgtactgggtgaagcagaggcctggacaaggccctgaatggattggagagattaatcctaccaatggtggtactaagttcaatgagaagttcaagagcaaggccacactgactgtagacaaatcctccagcacagcatatatacaactcagcagccttacatctgaggactctgcggtctattactgttcaagctcgggatatgattacgacgggagggcctactttgactactggggccagggcaccactctcacagtctcctca
SEQ ID NO:10
gacattgtgatgtcacagtctcctgcttccttagctgtatctctggggcagagggccaccatctcatacagggccagcaaaagtgtcagtacatctggctatagttatatgcactggaaccaacagaaaccaggacagccacccagactcctcatctatcttgtatccaacctagaatctggggtccctgccaggttcagtggcagtgggtctgggacagacttcaccctcaacatccatcctgtggaggaggaggatgctgcaacctattactgtcagcacattagggagcttacacgttcggaggggggaccaagctggaaa
将序列转化为氨基酸序列后，得到单克隆抗体40C10的重链可变区氨基酸序列SEQID NO:7和轻链可变区氨基酸序列SEQ ID NO:8。SEQ ID NO:7和SEQ ID NO:8的序列如下所示：
SEQ ID NO:7
Glu Val Gln Leu Gln Gln Ser GlyAla Glu Leu Val Lys Pro Gly Ala SerVal Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser His Tyr Met Tyr TrpVal Lys Gln Arg Pro Gly Gln Gly Pro Glu Trp Ile Gly Glu Ile Asn Pro Thr AsnGly Gly Thr Lys Phe Asn Glu Lys Phe Lys Ser Lys Ala Thr Leu Thr Val Asp LysSer Ser Ser ThrAla Tyr Ile Gln Leu Ser Ser Leu Thr Ser GluAsp SerAla Val TyrTyr Cys Ser Ser Ser Gly TyrAsp TyrAsp Gly Arg Ala Tyr Phe Asp Tyr Trp Gly GlnGlyThrThr Leu ThrVal Ser Ser
SEQ ID NO:8
Asp Ile Val Met Ser Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly GlnArg Ala Thr Ile Ser TyrArgAla Ser Lys SerVal Ser Thr Ser Gly Tyr Ser Tyr MetHis Trp Asn Gln Gln Lys Pro Gly Gln Pro Pro Arg Leu Leu Ile Tyr Leu Val SerAsn Leu Glu Ser Gly Val Pro AlaArg Phe Ser Gly Ser Gly Ser Gly ThrAsp Phe ThrLeu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln His IleArg Glu Leu ThrArg Ser Glu Gly GlyPro SerTrp Lys
通过分析，重链可变区CDR1、CDR2和CDR3的序列分别如SEQ ID NO:1、SEQ ID NO:2和SEQ ID NO:3所示，轻链可变区CDR1、CDR2和CDR3的序列分别如SEQ ID NO:4、SEQ ID NO:5和SEQ ID NO:6所示。
SEQ ID NO:1Ser His TyrMet Tyr
SEQ ID NO:2Glu Ile Asn Pro ThrAsn Gly Gly Thr Lys Phe Asn Glu Lys PheLys Ser
SEQ ID NO:3Ser Gly TyrAsp TyrAsp GlyArgAlaTyrPheAsp Tyr
SEQ ID NO:4Arg Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Tyr MetHis
SEQ ID NO:5LeuVal SerAsn Leu Glu Ser
SEQ ID NO:6Gln His IleArg
经测定，单克隆抗体40C10属于κ型的Ig抗体。
2.单克隆抗体40C10对于SFTSV抗原Gn的活性鉴定
2.1 SFTSV-Gn(20-452)蛋白真核表达抗原制备
为了鉴定筛选出的单克隆抗体40C10识别SFTSV的抗原，利用杆状病毒表达系统，构建SFTSV-Gn蛋白20到452位氨基酸到pFast-Bac-Dual的载体Polyhedrin启动子下，并在Gn蛋白质的C端加上Strep标签。利用Cellfectin转染试剂将pFBD-SFTSV-Gn(20-452)质粒转染到Sf9细胞中，3-5天后收集细胞上清，获得重组的杆状病毒vAc-SFTSV-Gn(20-452)。利用重组的杆状病毒感染Sf9细胞，3天后，收集感染后的细胞用于抗体的蛋白免疫印迹分析和间接免疫荧光验证。
2.2单克隆抗体40C10的蛋白免疫印迹分析
将本实施例2.1中的vAc-SFTSV-Gn感染的细胞进行蛋白免疫印迹分析，其中一组的vAc-SFTSV-Gn感染的细胞作为阳性对照，健康Sf9细胞作为空白对照(NC)，一抗使用SFTSV-GnR(稀释比1:1000)的兔多抗，分析结果如图4a所示，证明SFTSV-Gn(20-452)蛋白表达成功。另一组的同样的抗原，一抗使用单克隆抗体40C10，分析结果如图4b所示，证明单克隆抗体40C10不能识别SFTSV-Gn(20-452)蛋白的线性表位。
2.3单克隆抗体40C10的间接免疫荧光检测
将本实施例2.1中的vAc-SFTSV-Gn感染的细胞进行间接免疫荧光检测，其中试验组(Anti-40C10)采用单克隆抗体40C10作为一抗，阳性对照组(PC)采用SFTSV-GnR(稀释比1:1000)的兔多抗作为一抗，阴性对照组(NC)为健康的Sf9细胞。结果如图5所示，其中图5a、5b和5c分别为试验组、阳性对照组和阴性对照组的结果，证明单克隆抗体40C10能够识别SFTSV-Gn(20-452)蛋白的空间表位。
3.单克隆抗体40C10的体外中和活性评价
为评价实验室制备的单克隆抗体40C10对四个SFTSV毒株(WCH、HBMC5、HBGS13和NB24)和GTV的中和活性，进行病毒-抗体中和实验。首先，将100TCID50的病毒分别与0.8μg/mL、2.5μg/mL、7.4μg/mL、22.2μg/mL、66.6μg/mL和200μg/mL的单抗等体积混匀，在37℃孵育，1.5h后感染Vero细胞，3天后经免疫荧光检测评估抗体中和活性。中和效果如表2所示。
表2
结果表明，中和单抗40C10对SFTSV毒株(WCH、HBMC5、HBGS13和NB24)和GTV具有体外中和活性，抗体中和浓度分别为200μg/mL、66.7μg/mL、22.2μg/mL、66.7μg/mL和200μg/mL。说明40C10能够中和4种基因型的SFTSV毒株和GTV毒株。
4.单克隆抗体40C10小鼠体内保护性评价
选择三种基因型的SFTSV毒株(HBGS13-C2、HBMC5-C3、WCH-C4)进行中和抗体保护性研究。每只使用100LD50的毒株剂量腹腔注射6-8周鼠龄雌性A129小鼠(小鼠体重18-20g)，抗体按照每只老鼠30mg/kg的剂量在攻毒后的1h、24h、48h、72h注射单克隆抗体到小鼠腹腔内。连续12天内监测小鼠的体重和存活状况，分析体重变化和存活率。其中攻毒后的小鼠在相同时间点注射PBS作为对照组，每组小鼠各有6只。结果如图6所示，与对照组相比，40C10抗体能够保护A129小鼠，存活率为100％，同时小鼠体重范围为89-112％。说明40C10抗体能够在体内中和两种基因型的SFTSV毒株，并且保护小鼠，提高小鼠存活率。
通过以上可得，本发明的单克隆抗体40C10具有中和抗体活性，能够识别SFTSV-Gn蛋白的空间表位，不能识别线性表位。该抗体能够中和SFTSV的四种基因型的毒株(HBGS13-C2、HBMC5-C3、WCH-C4、NB24-J)和GTV病毒。并且能够在三种毒株(HBGS13-C2、HBMC5-C3、WCH-C4)感染A129小鼠模型上保护小鼠，具有广谱的中和效果。目前由SFTSV引起的发热伴血小板减少综合征疾病，没有特效药物和有效的疫苗。因此本发明的中和单克隆抗体40C10可以作为SFTSV抗病毒药物以及抗体用于SFTSV和GTV的防治。
以上所述仅为本发明的较佳实施例，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
序列表
<110> 中国科学院武汉病毒研究所
<120> 一种抗SFTSV的中和性单克隆抗体及其应用
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Ser His Tyr Met Tyr
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gaggttcagc tgcagcagtc tggggctgaa ctggtgaagc ctggggcttc agtgaagttg 60
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ggggtccctg ccaggttcag tggcagtggg tctgggacag acttcaccct caacatccat 240
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<212> DNA
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<212> DNA
<213> 人工序列(Artificial Sequence)
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<212> DNA
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<212> DNA
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<212> DNA
<213> 人工序列(Artificial Sequence)
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tgaggagacg gtgaccgtgg tccc 24
<210> 21
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
tgaggagact gtgagagtgg tgcc 24
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<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
tgcagagaca gtgaccagag tccc 24
<210> 23
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
tgaggagacg gtgactgagg ttcc 24
<210> 24
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
gacattgtga tgwcacagtc tcc 23
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<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
gatgttktga tgacccaaac tcc 23
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<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
gatattgtga tracbcaggc wgc 23
<210> 27
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
gacattgtgc tgacmcartc tcc 23
<210> 28
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
saaawtgtkc tcacccagtc tcc 23
<210> 29
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
gayatyvwga tgacmcagwc tcc 23
<210> 30
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
caaattgttc tcacccagtc tcc 23
<210> 31
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
tcattattgc aggtgcttgt ggg 23
<210> 32
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 32
tttgatttcc agcttggtgc ctcc 24
<210> 33
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
ttttatttcc agcttggtcc cccc 24
<210> 34
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
ttttatttcc agtctggtcc catc 24
<210> 35
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
ttttatttcc aactttgtcc ccga 24
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<211> 24
<212> DNA
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tttcagctcc agcttggtcc cagc 24
Claims (7)
1.一种抗SFTSV的中和性单克隆抗体，其特征在于，
所述单克隆抗体的重链可变区包含CDR1、CDR2和CDR3，重链可变区的CDR1、CDR2和CDR3的氨基酸序列分别如SEQ ID NO:1、SEQ ID NO:2和SEQ ID NO:3所示；
所述单克隆抗体的轻链可变区包含CDR1、CDR2和CDR3，轻链可变区的CDR1、CDR2和CDR3的氨基酸序列分别如SEQ ID NO:4、SEQ ID NO:5和SEQ ID NO:6所示。
2.根据权利要求1所述的抗SFTSV的中和性单克隆抗体，其特征在于，所述重链可变区的氨基酸序列如SEQ ID NO: 7所示，所述轻链可变区的氨基酸序列如SEQ ID NO:8所示。
3.根据权利要求2所述的抗SFTSV的中和性单克隆抗体，其特征在于，所述单克隆抗体的重链可变区由SEQ ID NO:9所示的核酸序列编码，所述单克隆抗体的轻链可变区由SEQID NO:10所示的核酸序列编码。
4.根据权利要求1或2所述的抗SFTSV的中和性单克隆抗体，其特征在于，所述重链的恒定区为IgG类型，所述轻链的恒定区为κ链。
5.权利要求1-4任一项所述的单克隆抗体在制备预防或治疗SFTSV感染的药物以及SFTSV和GTV感染的药物中的应用。
6.一种药物或疫苗，其特征在于，包括权利要求1-4任一项所述的单克隆抗体。
7.根据权利要求6所述的药物或疫苗，其特征在于，还包括药学上可接受的载剂、赋形剂、稀释剂、佐剂中的一种或多种。
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