CN115028715B - 一种抗新型冠状病毒的抗体或其抗原结合片段、试剂盒及应用 - Google Patents
一种抗新型冠状病毒的抗体或其抗原结合片段、试剂盒及应用 Download PDFInfo
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Abstract
本发明公开了一种抗新型冠状病毒的抗体或其抗原结合片段、试剂盒及应用，涉及生物医药高新技术领域。具有特定重链可变区和轻链可变区的抗体或其抗原结合片段能够特异性结合新型冠状病毒(SARS‑CoV‑2)的N蛋白，且具有较高的亲和力，该抗体针对型冠状病毒的N蛋白具有较高的灵敏度和特异性，本发明为新型冠状病毒的检测提供了更多以及性能更优的抗体选择。
Description
技术领域
本发明涉及生物医药高新技术领域，具体而言，涉及一种抗新型冠状病毒的抗体或其抗原结合片段、试剂盒及应用。
背景技术
新冠N蛋白也称新冠核衣壳蛋白，N蛋白在冠状病毒中含量丰富，是一种高度免疫原性蛋白，参与基因组复制N蛋白在和细胞信号通路调节。N蛋白是新冠检测和诊断非常重要的一个指标。
而受限于供人类诊断新冠的试剂较为稀缺，迫切的需要研发出更多的新冠诊断试剂。
鉴于此，特提出本发明。
发明内容
本发明的目的在于提供一种抗新型冠状病毒的抗体或其抗原结合片段、试剂盒及应用以开发相应的诊断试剂。
本发明是这样实现的：
本发明提供了一种抗新型冠状病毒的抗体或其抗原结合片段，其包括重链可变区(VH)和轻链可变区(VL)，VH包括以下的互补决定区：如SEQ ID NO.1所示的CDR-VH1、如SEQID NO.2所示的CDR-VH2和如SEQ ID NO.3所示的CDR-VH3，VL包括以下的互补决定区：如SEQID NO.4所示的CDR-VL1、如SEQ ID NO.5所示的 CDR-VL2和如SEQ ID NO.6所示的CDR-VL3。
发明人经过抗体筛选发现，具有上述重链可变区和轻链可变区的抗体能够特异性结合新型冠状病毒(SARS-CoV-2)的N蛋白，且具有较高的亲和力，该抗体针对型冠状病毒的N蛋白具有较高的灵敏度和特异性，本发明为新型冠状病毒的检测提供了更多以及性能更优的抗体选择。
在本发明应用较佳的实施方式中，上述抗体或其抗原结合片段与新型冠状病毒的N蛋白抗原以K≤1.86×109的亲和力结合。
在本发明应用较佳的实施方式中，上述抗体或其抗原结合片段还包括序列依次如SEQ ID NO：7-10所示的轻链骨架区FR1-L、FR2-L、 FR3-L及FR4-L，和/或，序列依次如SEQID NO:11-14所示的重链骨架区FR1-H、FR2-H、FR3-H及FR4-H。
需要说明的是，在其他的实施例中，本发明提供的抗体或其抗原结合片段的各骨架区氨基酸序列可以与上述对应骨架区(SEQ ID NO:7、8、9、10、11、12、13或14)可以具有至少80％、81％、82％、 83％、84％、85％、86％、87％、88％、89％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％的同源性。
在本发明应用较佳的实施方式中，上述抗原结合片段选自抗体的 F(ab’)2、Fab’、Fab、Fv和scFv中的任意一种。
在一种可选的实施方式中，抗体还包括恒定区；
在一种可选的实施方式中，恒定区的种属来源为小鼠。
上述抗体的抗原结合片段通常具有与其来源抗体相同的结合特异性。本领域技术人员根据本发明记载的内容容易理解到，上述抗体的功能性片段可以通过比如酶消化的方法(包括胃蛋白酶或木瓜蛋白酶)和/或通过化学还原分裂二硫键的方法获得。在本发明公开了完整抗体的结构基础上，本领域技术人员容易获得上述的抗原结合片段。
上述抗体的抗原结合片段还可以通过也是本领域技术人员所知的重组遗传学技术或通过例如自动肽合成仪，比如Applied BioSystems等销售的自动肽合成仪合成获得。
本发明还提供了一种检测新型冠状病毒的试剂或试剂盒，其包括抗新型冠状病毒的抗体或其抗原结合片段。
抗体或其功能性片段标记有可被检测的标记物。
在本发明应用较佳的实施方式中，上述可被检测的标记物选自荧光染料、催化底物显色的酶、放射性同位素、化学发光试剂和纳米颗粒类标记物。
在实际的使用过程中，本领域技术人员可以根据检测条件或实际需要选择合适的标记物，无论使用何种标记物，其均属于本发明的保护范围。
在本发明应用较佳的实施方式中，荧光染料包括但不限于荧光素类染料及其衍生物(例如包括但不限于异硫氰酸荧光素(FITC)羟基光素(FAM)、四氯光素(TET)等或其类似物)、罗丹明类染料及其衍生物(例如包括但不限于红色罗丹明(RBITC)、四甲基罗丹明(TAMRA)、罗丹明B(TRITC)等或其类似物)、Cy系列染料及其衍生物(例如包括但不限于Cy2、Cy3、Cy3B、Cy3.5、Cy5、Cy5.5、 Cy3等或其类似物)、Alexa系列染料及其衍生物(例如包括但不限于 AlexaFluor350、405、430、488、532、546、555、568、594、610、 33、647、680、700、750等或其类似物)和蛋白类染料及其衍生物 (例如包括但不限于藻红蛋白(PE)、藻蓝蛋白(PC)、别藻蓝蛋白 (APC)、多甲藻黄素-叶绿素蛋白(preCP)等)。
在本发明应用较佳的实施方式中，上述催化底物显色的酶选自辣根过氧化物酶、碱性磷酸酶、β-半乳糖苷酶、葡萄糖氧化酶、碳酸酐酶、乙酰胆碱酯酶以及6-磷酸葡萄糖脱氧酶。
在本发明应用较佳的实施方式中，上述放射性同位素选自212Bi、131I、111In、90Y、186Re、211At、125I、188Re、153Sm、213Bi、32P、94mTc、99mTc、203Pb、67Ga、68Ga、43Sc、47Sc、110mIn、97Ru、62Cu、64Cu、67Cu、68Cu、86Y、88Y、121Sn、161Tb、166Ho、105Rh、177Lu、172Lu和18F。
在本发明应用较佳的实施方式中，上述化学发光试剂选自鲁米诺及其衍生物、光泽精、甲壳动物荧光素及其衍生物、联吡啶钌及其衍生物、吖啶酯及其衍生物、二氧环乙烷及其衍生物、洛粉碱及其衍生物和过氧草酸盐及其衍生物。
在本发明应用较佳的实施方式中，上述纳米颗粒类标记物选自纳米颗粒和胶体。
在本发明应用较佳的实施方式中，上述纳米颗粒选自有机纳米颗粒、磁性纳米颗粒、量子点纳米颗粒和稀土络合物纳米颗粒。
在本发明应用较佳的实施方式中，上述胶体选自胶体金属、分散型染料、染料标记的微球、胶体硒和乳胶。
在本发明应用较佳的实施方式中，上述胶体金属选自胶体金和胶体银。
如上述抗新型冠状病毒的抗体或其抗原结合片段在制备检测新型冠状病毒的试剂或试剂盒中的应用。
在一种可选的实施方式中，试剂或试剂盒用于检测组织细胞中新型冠状病毒。
在一种可选的实施方式中，组织为被新冠感染后的肺脏。
本发明还提供了一种核酸分子，其编码上述的抗新型冠状病毒的抗体或其抗原结合片段。
如SEQ ID NO:15所示的序列编码上述抗体的重链可变区，如 SEQ ID NO:16所示的序列编码上述抗体的轻链可变区。
本发明还提供了一种表达盒或载体，其包括上述的核酸分子。
在一种可选的实施方式中，上述表达盒连接有用于调控上述核酸分子表达的调控序列，包括不限于启动子、增强子、信号肽编码序列、筛选标记基因、终止子、组氨酸标签等。
本发明还提供了一种重组菌或重组细胞，其含有上述的表达盒或载体。重组细胞可以是感受态细胞，例如选自大肠杆菌或酵母感受态细胞。
重组菌包括不限于大肠杆菌，例如BL21、DH5α、Top10等。
本发明还提供了一种制备上述的抗新型冠状病毒的抗体或其抗原结合片段的方法，其包括：培养上述的重组菌或重组细胞，从培养产物中分离纯化得到抗体或其抗原结合片段。
在本发明公开了抗体或其抗原结合片段的氨基酸序列的基础上，本领域技术人员容易想到采用基因工程技术或其他技术(化学合成、杂交瘤细胞)制备得到该抗体或其抗原结合片段，例如从能够重组表达如上任一项所述的抗体或其抗原结合片段的重组细胞的培养产物中分离纯化得到该抗体或其功能性片段，这对本领域技术人员来说是容易实现的，基于此，无论采用何种技术制备本发明的抗体或其抗原结合片段，其均属于本发明的保护范围。
本发明具有以下有益效果：
本发明提供的具有特定重链可变区和轻链可变区的抗体或其抗原结合片段能够特异性结合新型冠状病毒(SARS-CoV-2)的N蛋白，且具有较高的亲和力，该抗体针对型冠状病毒的N蛋白具有较高的灵敏度和特异性，本发明为新型冠状病毒的检测提供了更多以及性能更优的抗体选择。
具体实施方式
现将详细地提供本发明实施方式的参考，其一个或多个实例描述于下文。提供每一实例作为解释而非限制本发明。实际上，对本领域技术人员而言，显而易见的是，可以对本发明进行多种修改和变化而不背离本发明的范围或精神。例如，作为一个实施方式的部分而说明或描述的特征可以用于另一实施方式中，来产生更进一步的实施方式。
除非另外指明，否则实践本发明将采用细胞生物学、分子生物学 (包含重组技术)、微生物学、生物化学和免疫学的常规技术，所述常规技术在本领域技术人员的能力范围内。文献中充分解释了这种技术，如《分子克隆：实验室手册(Molecular Cloning:ALaboratory Manual)》，第二版(Sambrook等人，1989)；《寡核苷酸合成(OligonucleotideSynthesis)》(M.J.Gait编，1984)；《动物细胞培养 (Animal Cell Culture)》(R.I.Freshney编，1987)；《酶学方法(Methods in Enzymology)》(学术出版社有限公司(Academic Press，Inc.)；《实验免疫学手册(Handbook of Experimental Immunology)》(D.M.Weir和 C.C.Blackwell编)；《哺乳动物细胞用基因转移载体(Gene TransferVectors for Mammalian Cells)》(J.M.Miller和M.P.Calos编，1987)；《当代分子生物学方法(Current Protocols in Molecular Biology)》 (F.M.Ausubel等人编，1987)；《PCR:聚合酶链反应(PCR:The Polymerase Chain Reaction)》(Mullis等人编，1994)；以及《当代免疫学方法(Current Protocols in Immunology)》(J.E.Coligan等人编，1991)，所述文献中的每个文献均通过引用明确并入本文中。
为使本发明实施例的目的、技术方案和优点更加清楚，下面将对本发明实施例中的技术方案进行清楚、完整地描述。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市售购买获得的常规产品。
以下结合实施例对本发明的特征和性能作进一步的详细描述。
该抗体的生产包括小鼠免疫、细胞融合、抗体筛选、抗体纯化、抗体亲和力检测以及测序几个阶段。
实施例1
本实施例进行小鼠免疫。
1.制备抗原
首先设计N蛋白的引物序列，然后进行PCR获得目的基因，再将目的基因镶嵌在质粒上，再用电激的方法让质粒进入DH5α感受态细胞，然后将感受态细胞接种到含抗生素的平板上，放入37度烘箱摇菌，从而筛选出导入目的基因的大肠杆菌，再将大肠杆菌用超声破碎，然后用带his标签的镍柱过滤提纯，最后得到N蛋白，即抗原。
重组的人N蛋白序列如SEQ ID NO.17所示。
MGSSHHHHHHSSGLVPRGSHMSDNGPQNQRNAPRITFGGPS DSTGSNQNGERSGARSKQRRPQGLPNNTASWFTALTQHGKEDLK FPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANN AAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSRNSTPGSSRGTSPARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQG QTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQGNFG DQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKA DETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMSSADSTQA。
2.免疫小鼠
将制备好的N蛋白抗原和适量的PBS、福氏佐剂放入注射器中，注射器出水口用塞子塞住，放在乳化仪上搅拌充分乳化，使抗原佐剂形成稳定油包水的溶液。在初免和二免后的7-10天进行第一次尾血血清效价检测，经过2到4次的加强免疫得到良好的效价。选择血清效价高的免疫小鼠腹腔终免后进行后续的细胞融合。
实施例2
本实施例进行细胞融合。
细胞融合有多种方法，物理方法如电激，生物方法如灭活的病毒诱导，本实验选择的是用化学方法聚乙二醇诱导融合。
(1)复苏一管小鼠骨髓瘤细胞SP2/0，放在大皿培养至对数生长期。
(2)在融合前一天处死一只阴性鼠，在无菌操台用注射器吸取 6-8ml左右的HAT培养基，再吸入约2ml空气，注入小鼠腹腔，然后对小鼠腹部进行2-3分钟的按摩，再缓慢取出含滋养层细胞的HAT 培养基，然后将其铺板在96孔板，每孔100ul(滋养层细胞的作用就是为了增强杂交瘤细胞适应生存的能力)。
(3)到了第二天，先处死用N蛋白免疫免疫过的小鼠，然后采集眼球血留着作为后续ELISA的阳性对照。将老鼠放置在无菌操作台，取其脾脏，制成细胞悬液，将脾细胞悬液与SP2/0混匀(本次实验实验脾细胞和SP2/0细胞所选用培养均为RP1640)，用RP1640清洗两次,尽量弃尽上清。然后加入适量的PEG诱导脾脏B细胞和SP2/0 骨髓瘤细胞融合，再加入适量的RP1640终止融合。
(4)根据要铺板的数量，将融合后的细胞加入适量的HAT培养基，最后将融合细胞铺在滋养层细胞培养板上，每孔100ul(HAT培养基中含有叶酸类似物氨基蝶呤-A,可以阻断DNA的主要合成途径，因此细胞只能利用HGPRT和TK两种酶通过补救途径合成DNA。而骨髓瘤细胞不含该酶，因此无法合成DNA，导致慢慢死亡；B细胞虽然含有该酶，但是B细胞离开体外不能存活；而杂交瘤细胞既有该酶，又能在体外存活，所以最终只有杂交瘤细胞存活在融合孔中)。
实施例3
本实施例进行抗体筛选。
1.ELISA初筛:
7-10天后可在显微镜下观察存活的杂交瘤细胞生长情况，在铺板两周后，收集各孔上清，用ELISA方法用人N蛋白抗原进行杂交瘤细胞筛选。方法如下：(1)用100ul 2ug/ml人N蛋白抗原的PBS溶液包被酶标板37℃两小时。(2)PBST洗板3次后，加入150ul/孔的 3％脱脂奶粉的PBS溶液至于4℃封闭过夜。(3)再洗板三次，加入 80ul/孔的杂交瘤上清，37℃孵育1小时。(4)而后洗板三次。以100ul/ 孔加入1：7000稀释的辣根过氧化物酶标记的羊抗小鼠二抗，37℃孵育45分钟。(5)洗板3次，拍干。添加100ul/孔的TMB显色液，室温显色5-10分钟。(6)用50ul/孔，浓度为2M硫酸溶液终止。(7) 测定450纳米处各孔的吸光值。挑选出阳性的杂交瘤细胞。
2.第一轮单克隆培养：
选择ELISA为强阳性的融合孔来进行单克隆培养(有限稀释法)。首先挑选一个阳性孔扩配到24孔板，然后吸取10ul加到细胞计数板中进行计数，计算出细胞计数板四个区域细胞平均值，每个区域体积为10-4ml，从而得出细胞浓度，在算出100个细胞所需要的体积，取该体积量的液体加入10ml-HT培养中，然后10ml培养基平均加入96 孔板，这样就使得每孔大约一个细胞，进行第一轮单克隆培养，简称一克。
3.第二轮单克隆培养：
大概一周后，将一克孔上清取出做ELISA，最后挑出ELISA为强阳性的一克孔，再次利用有限稀释法(同上)进行第二轮单克隆培养。
实施例4
本实施例进行抗体纯化。
将第二轮单克隆培养后的细胞上清再次做ELISA进行阳性筛选。通过筛选后获得强阳性的单孔细胞株(单克隆细胞株N-11)，将单克隆细胞株进行扩培，将收集到的约1x106个细胞注射进挑选好的老鼠 (老鼠需提前一周在腹腔注射石蜡油)，再经过7-10天的等待，小鼠产生腹水，收集腹水后，用ProG柱子进行抗体纯化。提纯后，获得特异性抗N蛋白的小鼠单克隆抗体。再将纯化后的抗体用微量检测仪测其浓度，然后再将抗体保存。
实施例5
本实施例进行抗体亲和力检测。
将N蛋白抗原按照3mg/L、1.5mg/L、0.75mg/L、0.375mg/L包板；调整抗体浓度至10- 7mol/L水平，然后倍比稀释1:2～1:256，加至不同抗原包被量的孔中；加入二抗，TMB显色，检测450nm吸光值。根据抗原抗体结合S曲线，求出不同抗原浓度下，半数吸光值的抗体浓度。半数吸光值的抗体浓度可通过IGOR软件计算获得。这样就有四个抗体浓度(mol/L)，AB1，AB2，AB3，AB4。代入公式 K＝(N-1)/(N*AB’-AB)计算亲和力常数。AB代表抗原浓度为AG时，产生半数吸光值的抗体浓度；AB’代表抗原浓度为AG’时，产生半数吸光值的抗体浓度。N＝AG/AG’(AG>AG’)。当N＝2时，可以得到3个K值；当N＝4时，可以得到2个K值；当N＝8时，可以得到 1个K值。求出6个K值的平均值即为最终结果。
根据ELISA在OD450的检测结果，计算出各抗原包被浓度下的半数吸光值抗N蛋白抗体浓度，结果如下：
| 抗原包被浓度(AG)(ug/m) | 半数吸光值抗体浓度(AB)(10-9mol/L) |
| 3 | 2.6819 |
| 1.5 | 1.6094 |
| 0.75 | 4.2318 |
| 0.3725 | 18.308 |
由表可得6个K值平均值为3.9*108，即抗体亲和力常数为 K＝3.9*108。
实施例6
本实施例对抗体进行测序。
1.总RNA提取
使用Trizol试剂提取含有抗N蛋白抗体的小鼠单克隆细胞株 (N-11)中的总RNA。收集在9cm皿中培养的细胞到1.5ml离心管中，将上清吸干。加入1ml的Trizol试剂并将细胞吹打均匀使细胞裂解，将裂解后样品或匀浆液室温放置5-10min，使得核蛋白与核酸完全分离。加入0.2ml氯仿，剧烈振荡15sec，室温放置3min。12000rpm 4℃离心10min。吸取上层水相转移至干净的离心管中，加入等体积异丙醇，混匀，室温放置20min。12000rpm 4℃离心10min，弃上清。加入1ml 75％乙醇洗涤沉淀。12000rpm 4℃离心3min，弃上清。室温干燥5-10min。加入30-50ul RNase-free ddH2O。将所得到的RNA溶液置于-70℃保存或用于后续实验。
2.总RNA逆转录为cDNA
用超微量分光光度计测量RNA浓度，测完浓度后，取6ul的总 RNA+1ul Oligo dT+4ul RNase-free水(共11ul)。轻轻混匀后离心 3-5s，反应混合物在65℃温浴5mi预变性后，冰浴30s，然后离心3-5s，接着冰浴2min。在冰浴状态下加入4ul的5x缓冲液+1ul的dNTP混合物+1ul的RNase抑制剂+1ul逆转录酶(总共20ul体系)，轻轻混匀后离心3-5s，在PCR仪上42℃50min，85℃5min，完成cDNA 合成。随机引物适合于500bp以下的短链cDNA的合成，所转录的 RNA模板无需poly(A)尾，可转录5`端区域。
3.对表达该单克隆抗体轻重链的CDNA进行PCR扩增。
对于扩增抗体轻链可变区序列，配置PCR反应体系：2x Taq酶缓冲液25ul+FP-VL1ul+RP-VL 1ul+cDNA 2ul+ddH2O 21ul。对于扩增抗体重链可变区序列，配置PCR反应体系：2x Taq酶缓冲液25ul+FP-VH 1ul+RP-VH 1ul+cDNA 2ul+ddH20 21ul。重链和轻链可变区PCR扩增的温度循环如下(其中步骤2到4，重复35个循环)：
步骤1-预变性94℃，4min；
步骤2-变性94℃，30s；
步骤3-退火55℃，45s；
步骤4-延伸72℃，60s；
步骤5-72℃，10min；
步骤6-保存4℃。
4.测序
PCR产物经1％琼脂糖凝胶电泳分析，切出对应大小的DNA区段条带(VH的约为375bp，VL的约为325bp)，用SanPrep柱式DNA 胶回收试剂盒进行DNA提取。简述如下：从琼脂糖凝胶中割下含目标片段的胶块，称重；加入胶块重量3-6倍的buffer B2,50℃水浴 5-10min溶胶；将溶胶液移入吸附柱中，8000g离心30s；倒掉收集管中的液体；柱中加入500ulwash Solution，9000g离心30s，倒掉收集管中液体；再重复加wash solution一次，倒掉液体；孔吸附柱于9000g 离心1min；将吸附柱放入一个干净的1.5ml离心管中，在吸附膜中央加入15-40ul Elution Buffer,室温静置1min后，离心1min。获得制备的DNA溶液，纯化PCR产物经过测序获得抗体的可变区序列。测序结果见下表。
由单克隆细胞株N-11分泌的鼠抗人N蛋白单克隆抗体，其VH 和VL的核酸序列和对应氨基酸序列见下表：
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。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。
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Pro Thr Glu Pro Lys Lys Asp Lys Lys Lys Lys Ala Asp Glu Thr Gln
385 390 395 400
Ala Leu Pro Gln Arg Gln Lys Lys Gln Gln Thr Val Thr Leu Leu Pro
405 410 415
Ala Ala Asp Leu Asp Asp Phe Ser Lys Gln Leu Gln Gln Ser Met Ser
420 425 430
Ser Ala Asp Ser Thr Gln Ala
435
Claims (13)
1.一种抗新型冠状病毒的抗体或其抗原结合片段，其特征在于，其包括重链可变区（VH）和轻链可变区（VL），所述VH包括以下的互补决定区：如SEQ ID NO.1所示的CDR-VH1、如SEQ ID NO.2所示的CDR-VH2和如SEQ ID NO.3所示的CDR-VH3，所述VL包括以下的互补决定区：如SEQ ID NO.4所示的CDR-VL1、如SEQ ID NO.5所示的CDR-VL2和如SEQ ID NO.6所示的CDR-VL3；所述抗原结合片段选自所述抗体的F(ab’)2、Fab’、Fab、Fv和scFv中的任意一种。
2.根据权利要求1所述的抗新型冠状病毒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与新型冠状病毒的N蛋白抗原以K≤1.86×109的亲和力结合。
3.根据权利要求1或2所述的抗新型冠状病毒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还包括序列依次如SEQ ID NO：7-10所示的轻链骨架区FR1-L、FR2-L、FR3-L及FR4-L，和/或，序列依次如SEQ ID NO:11-14所示的重链骨架区FR1-H、FR2-H、FR3-H及FR4-H。
4.根据权利要求1-3任一项所述的抗新型冠状病毒的抗体或其抗原结合片段，其特征在于，所述抗体还包括恒定区。
5.根据权利要求4所述的抗新型冠状病毒的抗体或其抗原结合片段，其特征在于，所述恒定区的种属来源为小鼠。
6.一种检测新型冠状病毒的试剂或试剂盒，其特征在于，其包括如权利要求1-5任一项所述的抗新型冠状病毒的抗体或其抗原结合片段。
7.如权利要求1-5任一项所述的抗新型冠状病毒的抗体或其抗原结合片段在制备检测新型冠状病毒的试剂或试剂盒中的应用。
8.根据权利要求7所述的应用，其特征在于，所述试剂或试剂盒用于检测组织细胞中新型冠状病毒。
9.根据权利要求8所述的应用，其特征在于，所述组织为被新冠病毒感染过的肺脏。
10.一种核酸分子，其特征在于，其编码权利要求1-5任一项所述的抗新型冠状病毒的抗体或其抗原结合片段。
11.一种表达盒或载体，其特征在于，其包括权利要求10所述的核酸分子。
12.一种重组菌或重组细胞，其特征在于，其含有权利要求11所述的表达盒或载体。
13.一种制备如权利要求1-5任一项所述的抗新型冠状病毒的抗体或其抗原结合片段的方法，其特征在于，其包括：培养权利要求12所述的重组菌或重组细胞，从培养产物中分离纯化得到所述抗体或其抗原结合片段。
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Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008060331A2 (en) * | 2006-05-19 | 2008-05-22 | Amgen Inc. | Antibodies to sars coronavirus |
| WO2020036403A1 (ko) * | 2018-08-17 | 2020-02-20 | 한림대학교 산학협력단 | 메르스 코로나바이러스 s 단백질에 대한 단클론 항체 및 그 용도 |
| CN111592594A (zh) * | 2020-03-13 | 2020-08-28 | 北京大学 | 一种抗新型冠状病毒的单克隆抗体及其应用 |
| CN112239501A (zh) * | 2020-10-29 | 2021-01-19 | 东莞市朋志生物科技有限公司 | 抗新型冠状病毒的抗体、检测新型冠状病毒试剂和试剂盒 |
| CN112390879A (zh) * | 2021-01-21 | 2021-02-23 | 上海科技大学 | 靶向SARS-CoV-2的抗体及其制备方法和应用 |
| EP3800473A1 (en) * | 2020-03-25 | 2021-04-07 | National University of Singapore | Detection of antibodies to sarsr-cov |
| CN112979794A (zh) * | 2020-05-27 | 2021-06-18 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | 检测新型冠状病毒抗原的产品及其包含的抗体 |
| CN113249337A (zh) * | 2021-07-15 | 2021-08-13 | 天津一瑞生物科技股份有限公司 | 鼠抗新型冠状病毒n蛋白杂交瘤细胞株，单克隆抗体及应用 |
| CN113354729A (zh) * | 2020-03-06 | 2021-09-07 | 深圳市第三人民医院 | 一种抗新型冠状病毒的单克隆抗体及其应用 |
| CN113388030A (zh) * | 2021-08-17 | 2021-09-14 | 上海浙江大学高等研究院 | 单克隆抗体32c7及其制备方法和用途 |
| WO2021237534A1 (zh) * | 2020-05-27 | 2021-12-02 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | 抗新型冠状病毒的抗体、抗体组合、制备方法、包含其的检测试剂盒 |
| WO2021239949A1 (en) * | 2020-05-29 | 2021-12-02 | Deutsches Zentrum Für Neurodegenerative Erkrankungen E. V. (Dzne) | Human recombinant monoclonal antibody against sars-cov-2 spike glycoprotein |
-
2022
- 2022-06-23 CN CN202210723219.9A patent/CN115028715B/zh active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008060331A2 (en) * | 2006-05-19 | 2008-05-22 | Amgen Inc. | Antibodies to sars coronavirus |
| WO2020036403A1 (ko) * | 2018-08-17 | 2020-02-20 | 한림대학교 산학협력단 | 메르스 코로나바이러스 s 단백질에 대한 단클론 항체 및 그 용도 |
| CN113354729A (zh) * | 2020-03-06 | 2021-09-07 | 深圳市第三人民医院 | 一种抗新型冠状病毒的单克隆抗体及其应用 |
| CN111592594A (zh) * | 2020-03-13 | 2020-08-28 | 北京大学 | 一种抗新型冠状病毒的单克隆抗体及其应用 |
| EP3800473A1 (en) * | 2020-03-25 | 2021-04-07 | National University of Singapore | Detection of antibodies to sarsr-cov |
| CN112979794A (zh) * | 2020-05-27 | 2021-06-18 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | 检测新型冠状病毒抗原的产品及其包含的抗体 |
| WO2021237534A1 (zh) * | 2020-05-27 | 2021-12-02 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | 抗新型冠状病毒的抗体、抗体组合、制备方法、包含其的检测试剂盒 |
| WO2021239949A1 (en) * | 2020-05-29 | 2021-12-02 | Deutsches Zentrum Für Neurodegenerative Erkrankungen E. V. (Dzne) | Human recombinant monoclonal antibody against sars-cov-2 spike glycoprotein |
| CN112239501A (zh) * | 2020-10-29 | 2021-01-19 | 东莞市朋志生物科技有限公司 | 抗新型冠状病毒的抗体、检测新型冠状病毒试剂和试剂盒 |
| CN112390879A (zh) * | 2021-01-21 | 2021-02-23 | 上海科技大学 | 靶向SARS-CoV-2的抗体及其制备方法和应用 |
| CN113249337A (zh) * | 2021-07-15 | 2021-08-13 | 天津一瑞生物科技股份有限公司 | 鼠抗新型冠状病毒n蛋白杂交瘤细胞株，单克隆抗体及应用 |
| CN113388030A (zh) * | 2021-08-17 | 2021-09-14 | 上海浙江大学高等研究院 | 单克隆抗体32c7及其制备方法和用途 |
Non-Patent Citations (1)
| Title |
|---|
| anti-fluorescein monoclonal IgM heavy chain, partial [Mus musculus];NCBI;NCBI;1-4 * |
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