CN114634556B - 一种新冠病毒Delta和Omicron变异株嵌合抗原、其制备方法和应用 - Google Patents
一种新冠病毒Delta和Omicron变异株嵌合抗原、其制备方法和应用 Download PDFInfo
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Abstract
本发明涉及一种新型冠状病毒Delta和Omicron变异株的嵌合抗原、其制备方法和应用。本发明所述重组抗原由(1)来自新冠病毒Delta变异株的RBD蛋白的特定氨基酸序列和(2)来自新冠病毒Omicron变异株的RBD蛋白的特定氨基酸序列通过适当的连接序列或者直接串联而成。相较于新冠病毒原始毒株或其变异株的RBD同源二聚体，本发明的重组抗原能够更高效地激活广谱保护性抗体，对原始毒株以及当前的各种变异株都能起到很好的预防或治疗效果。
Description
技术领域
本发明涉及生物医药领域，具体涉及一种新型冠状病毒Delta和Omicron变异株嵌合抗原、其制备方法和应用。
背景技术
新型冠状病毒肺炎(也称COVID-19)是由新型冠状病毒(也称新冠病毒，SARS-CoV-2)感染导致的一种急性呼吸道传染病。新冠病毒属于冠状病毒科β-冠状病毒属，具有囊膜，是正链RNA病毒。新冠病毒表面的刺突(S)蛋白负责病毒的受体识别和膜融合，S蛋白上存在受体结合结构域(RBD)是一个重要的疫苗靶点，它激发中和抗体的产生，具有免疫聚焦的优势。在新冠疫情初期我们紧急攻关开发了基于新冠病毒RBD二聚体的重组亚单位蛋白疫苗ZF2001，在之后的临床实验中显示出良好的免疫原性和保护效果。
目前，新冠肺炎疫情在全球范围内仍很严峻，新冠病毒变异株不断出现和流行，其中有些会逃逸现有疫苗的免疫反应，引起突破性感染。特别是德尔塔(Delta)和奥密克戎(Omicron)变异株依次席卷全球，成为了优势流行毒株。Omicron变异株的S蛋白突变位点多达32处，对新冠病毒中和抗体药物和疫苗激活的体液免疫反应都有严重的免疫逃逸，对当前的疫情防控带来了严峻挑战。然而，有研究报道，以Omicron序列开发的疫苗激活的免疫反应虽然对Omicron变异株较强，但是对原型毒株以及其它毒株的交叉反应很弱，不适应当前原型株和多种变异株共存，且流行变异株仍在较快变化的情况，因此，需要研发一款针对当前的全球流行株均具有较强保护效果，可以诱导广谱的免疫反应的疫苗，这对于新冠疫情的防控可以起到至关重要的作用。
公开于该背景技术部分的信息仅仅旨在增加对本发明的总体背景的理解，而不应当被视为承认或以任何形式暗示该信息构成已为本领域一般技术人员所公知的现有技术。
发明内容
发明目的
本发明的目的在于提供一种新型冠状病毒Delta和Omicron变异株的重组嵌合抗原、其相关产品及其制备方法和应用。根据本发明的重组抗原为由(1)来自新型冠状病毒Delta变异株的RBD蛋白的特定氨基酸序列或与其具有至少90％，92％，95％，96％，97％，98％或99％同一性的氨基酸序列和(2)来自新型冠状病毒Omicron变异株的RBD蛋白的特定氨基酸序列或与其具有至少90％，92％，95％，96％，97％，98％或99％同一性的氨基酸序列直接串联或者通过适当的连接序列串联而形成的单链二聚体，其能够高效地激活广谱保护性抗体，对原始毒株以及当前的各种变异株都能起到很好的预防或治疗效果。
解决方案
为实现本发明目的，本发明提供了以下技术方案：
第一方面，本发明提供了一种新型冠状病毒Delta和Omicron变异株的重组嵌合抗原，所述重组抗原的氨基酸序列包括：按照(A-B)-(A-B’)或(A-B)-C-(A-B’)样式排列的氨基酸序列，其中：
A-B表示如SEQ ID NO:1所示的氨基酸序列或与其具有至少90％，92％，95％，96％，97％，98％或99％同一性的氨基酸序列，
A-B’表示如SEQ ID NO:2所示的氨基酸序列或与其具有至少90％，92％，95％，96％，97％，98％或99％同一性的氨基酸序列，并且
C表示连接序列。
在上述重组抗原的一种可行的实现方式中，C表示如(GGS)n所示的连接序列，其中，n表示GGS的个数，n为1至10之间的整数，优选为1-5之间的整数。
在上述重组抗原的一种优选的实施方案中，所述重组抗原的氨基酸序列包括：按照(A-B)-(A-B’)样式排列的氨基酸序列，其中：
A-B表示如SEQ ID NO:1所示的氨基酸序列或与其具有至少90％，92％，95％，96％，97％，98％或99％同一性的氨基酸序列，优选为如SEQ ID NO:1所示的氨基酸序列；
A-B’表示如SEQ ID NO:2所示的氨基酸序列或与其具有至少90％，92％，95％，96％，97％，98％或99％同一性的氨基酸序列，优选为如SEQ ID NO:2所示的氨基酸序列。
在一个优选的具体实施方案中，所述重组抗原的氨基酸序列如SEQ ID NO:3所示。
第二方面，本发明提供了一种如上述第一方面所述重组抗原的制备方法，其包括以下步骤：
在编码如上述第一方面所述重组抗原的核苷酸序列的5’端加上编码信号肽的序列，3’端加上组氨酸和终止密码子，进行克隆表达，筛选正确的重组子，然后将其转染表达系统的细胞进行表达，收集细胞培养上清，从中分离获得所述重组抗原。
在上述制备方法的一种可行的实现方式中，所述表达系统的细胞包括哺乳动物细胞、昆虫细胞、酵母细胞或细菌细胞；
可选地，所述哺乳动物细胞包括HEK293T细胞、HEK293F细胞、Expi293F细胞或CHO细胞；
可选地，所述细菌细胞包括大肠杆菌细胞。
第三方面，本发明提供了一种多核苷酸，其编码如上述第一方面所述的重组抗原。
所述多核苷酸为经人源密码子优化的核苷酸序列，可以为DNA或mRNA；
优选地，所述多核苷酸为如SEQ ID NO:4所示的核苷酸序列。
第四方面，本发明提供了一种核酸构建体，其包含如上述第三方面所述的多核苷酸，以及任选地，与所述多核苷酸可操作地连接的至少一个表达调控元件。
第五方面，本发明提供了一种表达载体，其包含如上述第四方面所述的核酸构建体。
第六方面，本发明提供了一种转化的细胞，其包括如上述第三方面所述的多核苷酸、如上述第四方面所述的核酸构建体或如上述第五方面所述的表达载体。
第七方面，本发明提供了如上述第一方面所述的重组抗原、如上述第三方面所述的多核苷酸、如上述第四方面所述的核酸构建体、如上述第五方面所述的表达载体或如上述第六方面所述的转化的细胞在制备新型冠状病毒疫苗中的应用。
第八方面，本发明提供了一种疫苗或免疫原性组合物，其包含如上述第一方面所述的重组抗原、如上述第三方面所述的多核苷酸、如上述第四方面所述的核酸构建体、如上述第五方面所述的表达载体或如上述第六方面所述的转化的细胞，以及生理学可接受的媒介物、佐剂、赋形剂、载体和/或稀释剂。
在一个优选的具体实施方案中，所述疫苗或免疫原性组合物为新型冠状病毒重组蛋白疫苗，其包括如上述第一方面所述的重组抗原和佐剂；
可选地，所述佐剂为选自以下佐剂中的一种或多种：铝佐剂、MF59佐剂和类MF59佐剂。
在另一个优选的具体实施方案中，所述疫苗或免疫原性组合物为新型冠状病毒DNA疫苗，其包括：
(1)真核表达载体；和
(2)构建入所述真核表达载体中的、编码如上述第一方面所述的重组抗原的DNA序列；
可选地，所述真核表达载体选自pGX0001、pVAX1、pCAGGS和pCDNA系列载体。
在另一个优选的具体实施方案中，所述疫苗或免疫原性组合物为新型冠状病毒mRNA疫苗，其包括编码如上述第一方面所述的重组抗原的mRNA序列和脂质纳米颗粒。
在另一个优选的具体实施方案中，所述疫苗或免疫原性组合物为新型冠状病毒-病毒载体疫苗，其包括：
(1)病毒骨架载体；和
(2)构建入所述病毒骨架载体中的、编码如上述第一方面所述的重组抗原的核酸序列；
可选地，所述病毒骨架载体选自以下病毒载体中的一种或几种：腺病毒载体、痘病毒载体、流感病毒载体、腺相关病毒载体。
在可行的实现方式中，所述疫苗或免疫原性组合物为鼻喷剂、口服制剂、栓剂或胃肠外制剂的形式；
优选地，所述鼻喷剂选自气雾剂、喷雾剂和粉雾剂；
优选地，所述口服制剂选自片剂、粉末剂、丸剂、散剂、颗粒剂、细粒剂、软/硬胶囊剂、薄膜包衣剂、小丸剂、舌下片和膏剂；
优选地，所述胃肠外制剂为经皮剂、软膏剂、硬膏剂、外用液剂、可注射或可推注制剂。
有益效果
本发明的发明人设计了一种新型冠状病毒Delta和Omicron变异株的重组嵌合抗原，该重组抗原由(1)来自新型冠状病毒Delta变异株的RBD蛋白的特定氨基酸序列或与其具有至少90％，92％，95％，96％，97％，98％或99％同一性的氨基酸序列和(2)来自新型冠状病毒Omicron变异株的RBD蛋白的特定氨基酸序列或与其具有至少90％，92％，95％，96％，97％，98％或99％同一性的氨基酸序列直接串联或者通过适当的连接序列串联而成，其可以诱导产生针对原始病毒株以及一系列主要变异毒株的高水平的中和抗体，有望成为预防新型冠状病毒的广谱疫苗。
附图说明
一个或多个实施例通过与之对应的附图中的图片进行示例性说明，这些示例性说明并不构成对实施例的限定。在这里专用的词“示例性”意为“用作例子、实施例或说明性”。这里作为“示例性”所说明的任何实施例不必解释为优于或好于其它实施例。
图1是本发明实施例1中构建和表达的新冠病毒原型毒株RBD二聚体(简称原型RBD-dimer)、Delta变异株RBD二聚体(简称Delta RBD-dimer)、Omicron变异株RBD二聚体(简称Omicron RBD-dimer)以及Delta RBD与Omicron RBD连接形成的嵌合RBD二聚体(简称Delta-Omicron嵌合RBD-dimer，代表本发明的重组抗原)的结构示意图。
图2是本发明实施例1中记载的，使用镍亲和柱纯化Delta-Omicron嵌合RBD-dimer蛋白的吸光度曲线，以及所收集洗脱峰的SDS-PAGE鉴定结果，箭头指示的位置是目的蛋白所在的洗脱峰。
图3是本发明实施例1中记载的，经过镍亲和柱纯化的、含Delta-Omicron嵌合RBD-dimer蛋白的洗脱液进行分子筛层析(以进一步纯化该蛋白)的吸光度曲线，以及所收集洗脱峰的SDS-PAGE鉴定结果(在非还原或还原条件下)，箭头指示的位置是目的蛋白所在的洗脱峰。
图4是本发明实施例1中记载的，经过镍亲和柱纯化的、含原型RBD-dimer蛋白的洗脱液进行分子筛层析(以进一步纯化该蛋白)的吸光度曲线，以及所收集洗脱峰的SDS-PAGE鉴定结果，箭头指示的位置是目的蛋白所在的洗脱峰。
图5是本发明实施例1中记载的，经过镍亲和柱纯化的、含Delta RBD-dimer蛋白的洗脱液进行分子筛层析(以进一步纯化该蛋白)的吸光度曲线，以及所收集洗脱峰的SDS-PAGE鉴定结果，箭头指示的位置是目的蛋白所在的洗脱峰。
图6是本发明实施例1中记载的，经过镍亲和柱纯化的、含Omicron RBD-dimer蛋白的洗脱液进行分子筛层析(以进一步纯化该蛋白)的吸光度曲线，以及所收集洗脱峰的SDS-PAGE鉴定结果，箭头指示的位置是目的蛋白所在的洗脱峰。
图7是本发明实施例2中记载的，新冠病毒原型毒株的RBD蛋白的不同视角的三维结构示意图，在其中，标记了Delta和Omicron变异株在RBD蛋白中的突变氨基酸位置、新冠病毒受体hACE2和5个代表性抗体(CB6、CV07-270、C110、S309和CR3022)的结合表位。
图8是本发明实施例2中记载的，通过表面等离子体共振实验检测的新冠病毒受体蛋白hACE2和5个不同抗体表位的代表性抗体CB6、CV07-270、C110、S309和CR3022与抗原蛋白的结合亲和力数据，以进行抗原蛋白的表位鉴定。
图9是本发明实施例3中记载的，通过分子筛层析技术纯化Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物的吸光度曲线，其中，一个洗脱峰是Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物，另一个洗脱峰是过量的CB6 Fab。
图10是本发明实施例3中记载的，Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物的冷冻电镜结构示意图。
图11是本发明实施例4中记载的，新冠病毒Alpha、Beta、Delta和Omicron变异株的S蛋白相对于新冠病毒原型毒株S蛋白的突变位点示意图。
图12是本发明实施例4中记载的，小鼠在免疫原第二次免疫后所采集的血清对新冠病毒原型毒株以及各个新冠病毒变异株的假病毒的中和滴度结果。
图13是本发明实施例5中记载的，对各免疫组小鼠进行Delta变异株攻毒后第3天采集的小鼠肺组织病毒载量的检测结果，其中，gRNA表示病毒基因组RNA，sgRNA表示病毒亚基因组RNA。
图14是本发明实施例5中记载的，免疫小鼠的血清对Delta变异株假病毒的中和抗体滴度与Delta变异株对该免疫小鼠攻毒后其肺组织的病毒gRNA载量的相关性分析。
图15是本发明实施例5中记载的，对各免疫组小鼠进行Omicron变异株攻毒后第3天采集的小鼠肺组织病毒载量的检测结果，其中，gRNA表示病毒基因组RNA，sgRNA表示病毒亚基因组RNA。
图16是本发明实施例5中记载的，免疫小鼠的血清对Omicron变异株假病毒的中和抗体滴度与Omicron变异株对该免疫小鼠攻毒后其肺组织的病毒gRNA载量的相关性分析。
图17是本发明实施例5中记载的，在用Delta或者Omicron变异株攻毒各组免疫小鼠后，每组小鼠的代表性肺组织HE染色病理图。
具体实施方式
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，显然，所描述的实施例是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
另外，为了更好的说明本发明，在下文的具体实施方式中给出了众多的具体细节。本领域技术人员应当理解，没有某些具体细节，本发明同样可以实施。在一些实施例中，对于本领域技术人员熟知的原料、元件、方法、手段等未作详细描述，以便于凸显本发明的主旨。
除非另有其它明确表示，否则在整个说明书和权利要求书中，术语“包括”或其变换如“包含”或“包括有”等等将被理解为包括所陈述的元件或组成部分，而并未排除其它元件或其它组成部分。
实施例1：SARS-CoV-2原型毒株RBD二聚体、Delta变异株RBD二聚体、Omicron变异株RBD二聚体以及Delta-Omicron嵌合RBD二聚体蛋白的构建、表达与纯化
按照图1所示的RBD二聚体结构示意图，分别设计新冠病毒原型毒株RBD二聚体(简称原型RBD-dimer)、Delta变异株RBD二聚体(简称Delta RBD-dimer)、Omicron变异株RBD二聚体(简称Omicron RBD-dimer)以及Delta RBD与Omicron RBD连接形成的嵌合RBD二聚体(简称Delta-Omicron嵌合RBD-dimer)的构建体，具体方案如下：
(1)将两个新冠病毒原型毒株的RBD序列(如SEQ ID NO:5所示)直接串联起来，在其N端连接信号肽(MIHSVFLLMFLLTPTES，SEQ ID NO.6)，在其C端加上6个组氨酸(HHHHHH)以及终止密码子，得到原型RBD-dimer构建体(其氨基酸序列如SEQ ID NO:7所示)；
(2)将两个新冠病毒Delta变异株的RBD序列(如SEQ ID NO:1所示)直接串联起来，在其N端连接信号肽(MIHSVFLLMFLLTPTES，SEQ ID NO.6)，在其C端加上6个组氨酸(HHHHHH)以及终止密码子，得到Delta RBD-dimer构建体(其氨基酸序列如SEQ ID NO:9所示)；
(3)将两个新冠病毒Omicron变异株的RBD序列(如SEQ ID NO:2所示)直接串联起来，在其N端连接信号肽(MIHSVFLLMFLLTPTES，SEQ ID NO.6)，在其C端加上6个组氨酸(HHHHHH)以及终止密码子，得到Omicron RBD-dimer构建体(其氨基酸序列如SEQ ID NO:11所示)；
(4)将新冠病毒Delta变异株的RBD序列(如SEQ ID NO:1所示)与Omicron变异株的RBD序列(如SEQ ID NO:2所示)直接串联起来，在其N端连接信号肽(MIHSVFLLMFLLTPTES，SEQ ID NO.6)，在其C端加上6个组氨酸(HHHHHH)以及终止密码子，得到Delta-Omicron嵌合RBD-dimer构建体(其氨基酸序列如SEQ ID NO:13所示)。
质粒构建：
上述四种构建体的氨基酸序列使用人源密码子优化，对应的DNA编码序列分别如SEQ ID NO:8、SEQ ID NO:10、SEQ ID NO:12和SEQ ID NO:14所示；在这些DNA编码序列的上游，分别加上Kozak序列gccacc，这四个包含Kozak序列的DNA序列由苏州金唯智生物科技有限公司合成；所合成的四段DNA序列通过EcoRI和XhoI酶切位点克隆到pCAGGS质粒，分别获得表达原型RBD-dimer、Delta RBD-dimer、Omicron RBD-dimer和Delta-Omicron嵌合RBD-dimer的表达质粒pCAGGS-原型、pCAGGS-Delta、pCAGGS-Omicron和pCAGGS-D-O嵌合。
蛋白表达与纯化：
将表达Delta-Omicron嵌合RBD-dimer蛋白的质粒转染Expi293FTM细胞，5天后收集上清，离心去除沉淀，再通过0.22μm的滤膜过滤，进一步除去杂质。将所得细胞上清在4℃通过镍亲和柱(His Trap,GE Healthcare)吸附，用缓冲液A(20mM Tris,150mM NaCl,pH 8.0)洗涤，除去非特异结合蛋白，然后用缓冲液B(20mM Tris,150mM NaCl,pH 8.0,300mM咪唑)将目的蛋白从His Trap上洗脱下来，收集洗脱峰处的洗脱液，用于目的蛋白的SDS-PAGE鉴定并用于后续的分子筛层析。Delta-Omicron嵌合RBD-dimer蛋白的镍亲和柱层析曲线以及其洗脱峰的SDS-PAGE鉴定结果见图2，图2左图中，箭头指向的位置是目的蛋白所在的洗脱峰。
然后，将上述收集的洗脱峰处的洗脱液用30kD浓缩管将洗脱液浓缩换液至分子筛层析缓冲液PBS(8mM Na2HPO4，136mM NaCl，2mM KH2PO4，2.6mM KCl，pH7.4)，终体积小于1ml；再通过SuperdexTM200 Increase 10/300GL柱子(GE Healthcare)进行分子筛层析，以进一步纯化目的蛋白，层析过程中，收集洗脱峰处的洗脱液，用于目的蛋白的SDS-PAGE鉴定。Delta-Omicron嵌合RBD-dimer蛋白的分子筛层析曲线及其洗脱峰的SDS-PAGE鉴定结果如图3所示，图3左图中，箭头指示的是目的蛋白对应的洗脱峰；此外，洗脱峰的SDS-PAGE凝胶电泳分析结果显示：洗脱蛋白的大小正确，证明了纯化得到了Delta-Omicron嵌合RBD-dimer蛋白，并且由电泳条带可知，所纯化的目的蛋白还具有较高的纯度和产量。
用同样的方法表达和纯化原型RBD-dimer、Delta RBD-dimer和Omicron RBD-dimer这三个蛋白，简言之，将它们的表达质粒分别转染Expi293FTM细胞，5天后收集上清，离心和过滤除去杂质。先通过镍亲和柱层析纯化，之后，原型RBD-dimer蛋白通过16/600/>200pg柱子(GE Healthcare)进行分子筛层析，Delta RBD-dimer和OmicronRBD-dimer这两个蛋白通过SuperdexTM200 Increase 10/300GL柱子(GE Healthcare)进行分子筛层析，以进一步纯化目的蛋白；原型RBD-dimer、Delta RBD-dimer和Omicron RBD-dimer的分子筛层析曲线及其洗脱峰的SDS-PAGE鉴定结果分别如图4、图5和图6所示，在各分子筛层析曲线上面，箭头指示的是目的蛋白对应的洗脱峰，将洗脱峰处的洗脱液进行SDS-PAGE凝胶电泳分析，结果显示：洗脱蛋白的大小均为60kD左右，其符合上述三种二聚体蛋白的分子大小，证明纯化得到了原型RBD-dimer、Delta RBD-dimer和Omicron RBD-dimer这三种二聚体蛋白，并且电泳条带单一，说明纯化后的蛋白具有较高的纯度，此外，原型RBD-dimer和Delta RBD-dimer还具有较高的产量。
实施例2：抗原表位鉴定和分析
发明人通过表面等离子共振技术(Surface Plasmon Resonance，SPR)鉴定抗原蛋白的RBD结合基序(RBM)以及主要的中和抗体表位的暴露，并检测抗原蛋白对新冠病毒的人受体——人血管紧张素转换酶(hACE2)的亲和力，以及对具有代表性的、靶向SARS-CoV-2RBD中的5个不同表位的单克隆抗体CB6(该抗体的具体信息请参见Ahuman neutralizingantibody targets the receptor-binding site of SARS-CoV-2.Nature,2020,PMID:32454512)、CV07-270(该抗体的具体信息请参见ATherapeutic Non-self-reactive SARS-CoV-2 Antibody Protects from Lung Pathology in a COVID-19 Hamster Model.Cell,2020,PMID:33058755)、C110(该抗体的具体信息请参见SARS-CoV-2 neutralizingantibody structures inform therapeutic strategies.Nature,2020,PMID:33045718)、S309(该抗体的具体信息请参见Cross-neutralization of SARS-CoV-2 by a humanmonoclonal SARS-CoV antibody.Nature,2020,PMID:32422645)和CR3022(该抗体的具体信息请参见Ahighly conserved cryptic epitope in the receptor binding domainsof SARS-CoV-2 and SARS-CoV.Science,2020,PMID:32245784)的Fab蛋白的亲和力；新冠病毒原型毒株的RBD蛋白中hACE2受体和5个单克隆抗体结合RBD的表位见图7。
亲和力测试实验中，测试了原型毒株的单体RBD蛋白、Delta变异株的单体RBD蛋白、Omicron变异株的单体RBD蛋白和Delta-Omicron嵌合RBD-dimer蛋白对hACE2受体和上述5个单克隆抗体的亲和力，并进行了对比分析。
亲和力测试方法如下：本测试是使用BIAcore8000(GE Healthcare)仪器进行操作的。测试前，将目的抗原蛋白换液到PBS-T缓冲液(10mM Na2HPO4，2mM KH2PO4,pH 7.4，137mMNaCl，2.7mM KCl，0.005％Tween 20)中。首先，利用氨基偶联的方法将抗原蛋白固定到CM5芯片上，目标响应值约为1000RU；然后，倍比稀释抗体Fab蛋白，稀释液作为流动相，以30μL/min的速度依次流过固定的抗原蛋白，得到不同的实时结合响应信号，收集的数据使用BIAevaluation Version 4.1(GE Healthcare)软件、按照1:1结合模型进行计算，最终得到抗原蛋白与抗体结合的亲和力。
亲和力测试结果如图8所示，由图8可知：所有的抗原蛋白都与hACE2受体具有相似的亲和力，KD值在5.09–8.44nM范围；在与上述五种单抗的亲和力方面，Delta RBD单体蛋白丧失了结合C110抗体的结合活性，Omicron RBD单体蛋白丧失了结合CB6抗体的结合活性，表明：Delta和Omicron变异株会逃逸一些原型毒株感染或者以原型毒株序列设计的疫苗诱导所产生的抗体反应；作为对比，Delta-Omicron嵌合RBD-dimer蛋白可以结合所有检测的代表性单抗，且Delta-Omicron嵌合RBD-dimer蛋白结合各个单抗的亲和力数值与Delta或Omicron单体RBD这两个蛋白结合各个单抗的亲和力中较强的一方相当，表明：Delta-Omicron嵌合RBD-dimer蛋白组合了Delta和Omicron的表位特征，且很好地暴露了受体结合位点以及展示了主要的中和抗体表位构象。
实施例3：本发明的Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物的电镜结构解析
将Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab蛋白混合，在4℃条件孵育12小时。然后通过SuperdexTM200 Increase 10/300GL柱子(GE Healthcare)进行分子筛层析(pH8.0)，以纯化Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab蛋白的复合物，其分子筛层析曲线如图9所示；此外，收集两个洗脱峰处的洗脱液，并对其进行SDS-PAGE鉴定，由SDS-PAGE鉴定结果(数据未示出)可知：图9的其中一个洗脱峰是Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物，另一个峰是过量的CB6 Fab，这说明：Delta-Omicron嵌合RBD-dimer蛋白可以与CB6 Fab结合并形成了复合物。
此外，将上述收集的Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物的洗脱液进行浓缩，浓缩后用于冷冻电镜分析，程序如下：
事先准备好制样用的Quantifoil载网(规格1.2/1.3)，进行辉光放电亲水化处理。随后将制备好的Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物滴加在上述准备好的载网上，用自动制样机器Vitrobot Mark IV将载网迅速插入液态乙烷中完成制样。
数据收集是用300kV Titan Krios透射电镜(Thermo Fisher公司)搭配K2直接电子探测器相机进行的，使用Serial-EM自动收集程序收集大量照片。然后，使用MotionCor2软件对收集到的原始数据进行漂移校正，用CTFFIND4.1软件对图片进行衬度传递函数校正，使用Relion-3.1软件对图片进一步处理和最后的三维重构。
本发明Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物的冷冻电镜图像如图10所示，由图10可见：在Delta-Omicron嵌合RBD-dimer蛋白与CB6 Fab的复合物中，DeltaRBD和Omicron RBD对称分布，呈“双肺状”；并且，Delta RBD可以结合CB6 Fab，Delta-Omicron嵌合RBD-dimer蛋白的主要表位完全暴露，有利于激活免疫反应。
实施例4：检测Delta-Omicron嵌合RBD-dimer蛋白诱导的体液免疫反应
为了检测本发明的Delta-Omicron嵌合RBD-dimer蛋白的免疫原性，我们将实施例1所得纯化的原型RBD-dimer、Delta RBD-dimer、Omicron RBD-dimer和Delta-Omicron嵌合RBD-dimer蛋白作为免疫原分别免疫BALB/c小鼠，阴性对照(Sham组)采用PBS溶液进行免疫，每组10只小鼠。所使用的BALB/c小鼠从维通利华公司购买，均为雌性，7-9周龄。小鼠分组及免疫剂量情况如表1所示。
表1新型冠状病毒RBD二聚体疫苗免疫小鼠分组及剂量
具体免疫程序如下：
将免疫原用PBS稀释至40μg/ml，将稀释后的免疫原与AddaVax佐剂(一种类似于MF59的疫苗佐剂)按照体积比1:1的比例混合、乳化，制备成疫苗。Sham组为PBS溶液与AddaVax佐剂混合，以制备疫苗对照。所得疫苗通过肌肉注射的方式对BALB/c小鼠进行免疫，所有小鼠分别在第0天、第21天进行第一次和第二次免疫，每次100μL的接种体积(含抗原蛋白2μg)。在第35天，对小鼠进行取血，离心收集血清，所得血清于-80℃冰箱保存。
使用新冠病毒假病毒，分别检测所收集的免疫小鼠血清对新冠病毒原型毒株和变异株的假病毒的50％假病毒中和滴度(pVNT50)，其中变异株包括Alpha、Beta、Delta、Omicron变异株，各变异株的S蛋白相对于原型毒株S蛋白的突变位点见图11。
本实施例中使用的新冠病毒假病毒为基于水疱性口炎病毒(VSV)骨架制备的展示新冠病毒S蛋白的假病毒，制备方法参见本课题组已经公开发表论文的方法部分(Effectsof a Prolonged Booster Interval on Neutralization of Omicron Variant,N Engl JMed,2022,PMID:35081296)。
检测新冠病毒假病毒(以下简称假病毒)中和抗体滴度的方法如下：在96孔板中，将免疫小鼠血清按2倍梯度倍比稀释，然后与假病毒分别混合，空白培养基也与假病毒混合作为对照，37℃孵育1小时。将免疫血清-假病毒混合液转移至已铺满Vero细胞的96孔板中。15小时后，通过CQ1共聚焦细胞成像仪(Yokogawa)计算阳性细胞数值，然后在GraphPadPrism软件中绘制拟合曲线，计算50％中和时对应的血清稀释度的倒数，即为50％假病毒中和滴度pVNT50。假病毒中和抗体滴度检测结果如图12所示。
由图12可见：
1)对于原型RBD-dimer，其免疫小鼠血清对原型毒株假病毒的中和抗体滴度的几何平均值(GMT)是3009，但是对部分变异株的假病毒的中和效果有所下降，包括Beta(GMT，1112)，Delta(GMT，2059)，Omicron(GMT，374)。
2)对于Delta RBD-dimer，其免疫小鼠血清对Delta假病毒的中和抗体滴度GMT是16722，但是对Beta假病毒的中和抗体滴度GMT是756，对Omicron假病毒的中和抗体滴度GMT是633，对Beta和Omicron变异株效果不佳。
3)对于Omicron RBD-dimer，其免疫小鼠血清对Omicron假病毒有一定的中和抗体活性，但是滴度较低(GMT＝43)，而且对原型毒株和Alpha、Beta、Delta变异株没有中和活性(低于检测限)。
4)对于Delta-Omicron嵌合RBD-dimer蛋白疫苗，其诱导了较为平衡的抗体反应，其免疫小鼠血清对假病毒的中和抗体滴度GMT分别是4518(原型)，5576(Alpha)，2263(Beta)，38387(Delta)和7194(Omicron)，对于这5个假病毒，Delta-Omicron嵌合RBD-dimer蛋白疫苗免疫的小鼠血清的中和抗体GMT均高于其它三个蛋白疫苗，显示出免疫原性强且广谱的优势。
实施例5：新冠病毒活病毒攻毒实验验证疫苗效果
为了进一步探索Delta-Omicron嵌合RBD-dimer蛋白疫苗的保护效果，对如上述实施例4所制备的Sham组、原型RBD-dimer免疫组和Delta-Omicron嵌合RBD-dimer免疫组小鼠分别进行SARS-CoV-2 Delta和Omicron变异株的活病毒攻毒实验。
对每个实验组中的5只小鼠进行新冠病毒Delta变异株的活病毒攻毒实验，对每个实验组中的另外5只小鼠进行新冠病毒Omicron变异株的活病毒攻毒实验。由于BALB/c小鼠对Delta变异株不易感，因此，采用如下的Delta变异株攻毒实验方法：采用滴鼻转导8×109vp表达受体蛋白hACE2的重组5型腺病毒(Ad5-hACE2)，建立瞬时表达hACE2的模型，在转导Ad5-hACE2五天后，经滴鼻感染6×105TCID50 Delta变异株(CCPM-B-V-049-2105-8)。此外，Omicron变异株的攻毒实验方法如下：小鼠直接通过滴鼻感染6×105TCID50新冠病毒Omicron变异株(BA.1,CCPM-B-V-049-2112-18)。
在新冠病毒Delta或Omicron变异株感染后第3天，对小鼠实施安乐死并进行解剖；每只小鼠取出肺，分成2份：一份匀浆研磨，提取病毒核酸，使用qRT-PCR方法对病毒的病毒基因组gRNA和亚基因组sgRNA进行定量，gRNA代表了全部病毒核酸，sgRNA代表正在复制过程的病毒核酸，是病毒复制水平的指标；另一份经多聚甲醛固定后，进行苏木精和伊红(H&E)染色染色，观察组织病理。
检测病毒gRNA和sgRNA的方法如下：将小鼠肺组织匀浆后，取组织匀浆上清液使用Direct-zol RNA MiniPrep试剂盒(Zymo Research公司，货号R2052)提取病毒RNA。在CFX384 Touch real-time PCR检测系统(Bio-Rad)上，使用TaqMan Fast Virus 1-StepMaster Mix试剂盒(Thermo Fisher Scientific，货号4444436)，进行SARS-CoV-2特异性定量逆转录PCR(qRT-PCR)检测。采用两组引物和探针分别检测SARS-CoV-2 Delta和Omicron病毒基因组gRNA，并采用一组引物和探针检测Delta和Omicron病毒sgRNA。
检测SARS-CoV-2 Delta gRNA的引物探针序列如下：
F,GACCCCAAAATCAGCGAAAT(SEQ ID NO：15)；
R,TCTGGTTACTGCCAGTTGAATCTG(SEQ ID NO：16)；
probe-Delta,FAM-ACCCCGCATTACGTTTGGTGGACC(SEQ ID NO：17)-BHQ1。
检测SARS-CoV-2 Omicron gRNA的引物序列同Delta，即(SEQ ID NO：15)和(SEQID NO：16),检测SARS-CoV-2 Omicron gRNA的探针序列如下：
probe-Omicron：FAM-ACTCCGCATTACGTTTGGTGGACC(SEQ ID NO：18)-BHQ1；
检测SARS-CoV-2 Delta和Omicron sgRNA的引物探针序列如下：
sgRNA-F,CGATCTCTTGTAGATCTGTTCTC(SEQ ID NO：19)；
sgRNA-R,ATATTGCAGCAGTACGCACACA(SEQ ID NO：20)；
sgRNA-probe,FAM-ACACTAGCCATCCTTACTGCGCTTCG(SEQ ID NO：21)-BHQ1。
Delta变异株的活病毒攻毒实验后，小鼠肺组织的病毒载量检测结果如图13所示；由图13可知，对于用新冠病毒Delta变异株攻毒的小鼠，对照组小鼠检测到高水平的gRNA(均值：1.09×1010拷贝/g肺组织)和sgRNA(均值：1.70×108拷贝/g肺组织)，相比之下，在疫苗免疫后的小鼠中检测到的病毒载量(包括gRNA和sgRNA)均显著降低，其中原型RBD-dimer和Delta-Omicron嵌合RBD-dimer免疫组的肺组织gRNA平均值分别为1.43×108拷贝/g和2.37×107拷贝/g肺组织。此外，Delta-Omicron嵌合RBD-dimer疫苗组所有小鼠均未检测到肺组织病毒sgRNA，表明它们完全抑制了病毒复制；而原型RBD-dimer组5只小鼠中有3只小鼠sgRNA呈阳性，原型RBD-dimer组滴度均值是1.07×106拷贝/g肺组织(图13)，这表明：相较于原型RBD-dimer，Delta-Omicron嵌合RBD-dimer对新冠病毒原型毒株的抑制效果明显更好。
将每只免疫小鼠的血清对新冠病毒Delta变异株假病毒的中和抗体滴度与攻毒后对应的肺组织病毒gRNA基于线性模型进行相关性分析，结果图14，由图14可以看到，中和抗体滴度与攻毒后肺组织中新冠病毒Delta变异株gRNA的相关性较高(r＝-0.8889，P<0.0001)，表明：中和抗体滴度越高，病毒的抑制效果越明显。
Omicron变异株的活病毒攻毒实验后，小鼠肺组织的病毒载量检测结果如图15所示；由图15可知，对于用新冠病毒Omicron变异株攻毒的小鼠，对照组小鼠检测到高水平的gRNA(均值：1.04×109拷贝/g肺组织)和sgRNA(均值：1.73×107拷贝/g肺组织)，相比之下，原型RBD-dimer免疫组和Delta-Omicron嵌合RBD-dimer免疫组的肺组织gRNA平均值分别为3.68×107拷贝/g和1.61×107拷贝/g肺组织。此外，Delta-Omicron嵌合RBD-dimer疫苗组的所有小鼠均未检测到肺组织病毒sgRNA，表明它们完全抑制了病毒复制，而原型RBD-dimer组的5只小鼠中有2只小鼠sgRNA呈阳性，原型RBD-dimer组滴度均值是2.41×104拷贝/g肺组织，表明：相较于原型RBD-dimer，Delta-Omicron嵌合RBD-dimer对新冠病毒Omicron变异株的抑制效果明显更好。
将每只小鼠的血清对新冠病毒Omicron变异株的假病毒的中和抗体滴度与新冠病毒Omicron变异株攻毒后对应的肺组织病毒gRNA基于线性模型进行相关性分析，结果图16，由图16可以看到，中和抗体滴度与攻毒后肺组织原型株新冠病毒gRNA的相关性较高(r＝-0.7362，P＝0.0017)，表明：中和抗体滴度越高，病毒的抑制效果越明显。
各实验组小鼠在用新冠病毒Delta变异株或Omicron变异株攻毒后的肺组织病理学结果如图17所示。通过分析图17中各实验组小鼠的肺组织病理学看出，对照组小鼠(Sham)在感染新冠病毒Delta或者Omicron变异株后，其肺部病理变化呈现出中度至重度的病变，包括肺泡腔消失、肺出血和弥漫性炎症细胞浸润；相比之下，接种原型RBD-dimer和Delta-Omicron嵌合RBD-dimer疫苗的小鼠只表现出轻微的肺损伤，显著减轻了肺炎(图17)。此外，小鼠的肺组织病理学结果表明：与原型RBD-dimer相比，Delta-Omicron嵌合RBD-dimer的保护效果更好(图17)，这些肺组织病理学结果与上述肺组织病毒gRNA测定的趋势一致，证明了Delta-Omicron嵌合RBD-dimer蛋白疫苗确实可以为新冠病毒不同毒株提供较为平衡且高效的保护作用，尤其是近期流行的Delta和Omicron变异株。
最后应说明的是：以上实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述实施例对本发明进行了详细的说明，本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明技术方案的精神和范围。
SEQUENCE LISTING
<110> 中国科学院微生物研究所
<120> 一种新冠病毒Delta和Omicron变异株嵌合抗原、其制备方法和应用
<130> 1087-220044F
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Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
260 265 270
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
275 280 285
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
290 295 300
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
305 310 315 320
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile
325 330 335
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
340 345 350
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
355 360 365
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
370 375 380
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly
385 390 395 400
Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
405 410 415
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
420 425 430
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
435 440 445
Thr Asn Leu Val Lys Asn Lys His His His His His His
450 455 460
<210> 8
<211> 1386
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的多核苷酸
<220>
<221> misc_feature
<222> (1)..(1386)
<223> 原型RBD-dimer构建体的核苷酸序列
<400> 8
atgatccact cagtgtttct cttaatgttt ctactaactc ccacggagtc gcgagtgcag 60
cctaccgaaa gcatcgtccg tttcccgaat attactaatc tctgtccatt cggagaagtc 120
ttcaatgcca cccgattcgc ttccgtttac gcgtggaacc gtaaacgaat atctaattgt 180
gttgcggact attccgtgtt gtacaactca gcatcattct ctacttttaa atgctatgga 240
gtgtcgccga ctaaactcaa cgacttgtgt ttcactaatg tttatgctga ctctttcgtt 300
attcgtggag acgaagttcg tcaaatcgca ccagggcaaa ctggcaagat tgcggactat 360
aattataagc tgccagatga ctttaccgga tgtgtaatag cctggaactc aaataatctc 420
gacagtaaag tgggaggcaa ctataattat ctttatcgac tcttcagaaa gtctaacctt 480
aagccatttg aacgtgacat ttctacagaa atttaccaag ccggctctac accttgcaat 540
ggcgtggaag ggtttaactg ttatttccca ttacagtctt atggtttcca gccaactaat 600
ggtgtgggat accaacctta ccgcgtcgtt gtcctgtcgt ttgaattgct tcacgcacca 660
gccaccgttt gtgggccaaa gaagagcact aatctcgtaa agaataaacg tgttcagcct 720
actgaatcga tcgtgaggtt cccaaatatt accaatctgt gtccgttcgg agaggtcttc 780
aatgcgactc gattcgcgtc tgtttacgcc tggaacagga aacggattag caattgtgtc 840
gctgactatt cggtcttata caactctgca tcattctcaa ccttcaagtg ttatggtgtc 900
agccctacaa agctgaatga cttatgtttc accaatgttt atgcggacag tttcgtaata 960
cgaggtgatg aagtccgcca aattgcaccc ggacaaaccg gcaagatagc cgactataat 1020
tataagctcc ctgatgactt tacgggctgt gtcatagctt ggaatagtaa taatttggac 1080
tcgaaagtgg gaggtaatta taattatctc tatagactgt tccggaaatc aaatctcaag 1140
ccctttgaac gggacataag tacagaaatc taccaagctg gttccacgcc gtgtaatgga 1200
gtcgaggggt ttaactgtta tttcccgctc cagtcgtatg ggttccagcc aacgaatggc 1260
gtcggatacc aaccttaccg cgttgtagta ttaagctttg aactgttgca cgcgcccgcg 1320
actgtttgtg gcccgaagaa gtcgactaat ctagtaaaga ataagcatca tcaccaccac 1380
cactga 1386
<210> 9
<211> 461
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成多肽
<220>
<221> MISC_FEATURE
<222> (1)..(461)
<223> Delta RBD-dimer构建体的氨基酸序列
<400> 9
Met Ile His Ser Val Phe Leu Leu Met Phe Leu Leu Thr Pro Thr Glu
1 5 10 15
Ser Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
20 25 30
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
35 40 45
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
50 55 60
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
65 70 75 80
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
85 90 95
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
100 105 110
Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
115 120 125
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
130 135 140
Gly Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu
145 150 155 160
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
165 170 175
Lys Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln
180 185 190
Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg
195 200 205
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
210 215 220
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
225 230 235 240
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
245 250 255
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
260 265 270
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
275 280 285
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
290 295 300
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
305 310 315 320
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile
325 330 335
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
340 345 350
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
355 360 365
Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
370 375 380
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Lys Pro Cys Asn Gly
385 390 395 400
Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
405 410 415
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
420 425 430
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
435 440 445
Thr Asn Leu Val Lys Asn Lys His His His His His His
450 455 460
<210> 10
<211> 1386
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的多核苷酸
<220>
<221> misc_feature
<222> (1)..(1386)
<223> Delta RBD-dimer构建体的核苷酸序列
<400> 10
atgatccaca gcgtgttcct gctgatgttc ctcctgaccc ccacagagag ccgggtgcaa 60
cccaccgaga gcatcgtgag attccccaac atcaccaacc tgtgtccctt cggggaagtg 120
tttaacgcca caagattcgc tagcgtgtac gcctggaata gaaagcggat cagcaattgc 180
gtggccgatt acagcgtgct gtacaattcc gctagcttca gcaccttcaa gtgttacggc 240
gtgagcccca ccaagctgaa tgacctctgc tttaccaatg tgtacgccga cagctttgtg 300
attagaggcg acgaagtgag acagatcgcc cctgggcaga ccggcaagat tgctgactat 360
aattacaagc tgcctgatga cttcaccggc tgtgtgattg cctggaacag caacaacctg 420
gattccaagg tggggggcaa ttacaactac cggtaccggc tgttcagaaa gtccaacctg 480
aagcccttcg aaagagacat ctccaccgag atttaccaag ccggcagcaa gccctgcaac 540
ggcgtggagg gcttcaactg ctattttccc ctgcaaagct atggctttca gcccaccaac 600
ggcgtgggct atcaacccta tagagtggtc gtgctgagct tcgagctgct gcacgccccc 660
gccaccgtct gtggccccaa gaagagcacc aacctggtca agaacaagag agtgcagccc 720
accgagagca tcgtgcggtt ccccaatatc accaacctct gccctttcgg cgaggtgttc 780
aacgctacaa gattcgcttc cgtgtacgcc tggaacagaa aacggatttc caactgcgtg 840
gccgactaca gcgtgctcta caacagcgct agcttctcca cattcaaatg ctacggcgtg 900
agccccacaa agctgaatga cctgtgcttc accaacgtgt atgccgacag cttcgtgatc 960
agaggcgacg aggtcagaca gatcgctccc gggcagaccg ggaagatcgc cgactacaac 1020
tataagctgc ccgacgattt taccggctgc gtgatcgcct ggaatagcaa caacctcgac 1080
agcaaggtgg gcggcaacta taattataga taccggctgt ttcggaagag caacctgaag 1140
ccctttgaga gagacatcag cacagagatc taccaagccg gcagcaaacc ctgtaacggc 1200
gtcgagggct ttaactgtta cttccccctg cagagctatg ggtttcagcc taccaatggc 1260
gtgggctatc agccctatag agtggtcgtg ctctccttcg aactgctgca cgctcctgcc 1320
accgtctgcg gccccaaaaa gtccacaaac ctggtgaaaa acaagcatca ccatcaccat 1380
cactga 1386
<210> 11
<211> 461
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成多肽
<220>
<221> MISC_FEATURE
<222> (1)..(461)
<223> Omicron RBD-dimer构建体的氨基酸序列
<400> 11
Met Ile His Ser Val Phe Leu Leu Met Phe Leu Leu Thr Pro Thr Glu
1 5 10 15
Ser Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
20 25 30
Asn Leu Cys Pro Phe Asp Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
35 40 45
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
50 55 60
Ser Val Leu Tyr Asn Leu Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly
65 70 75 80
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
85 90 95
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
100 105 110
Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
115 120 125
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Lys Leu Asp Ser Lys Val
130 135 140
Ser Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu
145 150 155 160
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn
165 170 175
Lys Pro Cys Asn Gly Val Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg
180 185 190
Ser Tyr Ser Phe Arg Pro Thr Tyr Gly Val Gly His Gln Pro Tyr Arg
195 200 205
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
210 215 220
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
225 230 235 240
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
245 250 255
Asp Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
260 265 270
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
275 280 285
Leu Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
290 295 300
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
305 310 315 320
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile
325 330 335
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
340 345 350
Ala Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser Gly Asn Tyr Asn
355 360 365
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
370 375 380
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys Pro Cys Asn Gly
385 390 395 400
Val Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg Ser Tyr Ser Phe Arg
405 410 415
Pro Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val Val Val Leu Ser
420 425 430
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
435 440 445
Thr Asn Leu Val Lys Asn Lys His His His His His His
450 455 460
<210> 12
<211> 1386
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的多核苷酸
<220>
<221> misc_feature
<222> (1)..(1386)
<223> Omicron RBD-dimer构建体的核苷酸序列
<400> 12
atgatccaca gcgtgttcct gctgatgttc ctcctgaccc ccacagagag ccgggtgcaa 60
cccaccgaga gcatcgtgag attccccaac atcaccaacc tgtgtccctt cgatgaagtg 120
tttaacgcca caagattcgc tagcgtgtac gcctggaata gaaagcggat cagcaattgc 180
gtggccgatt acagcgtgct gtacaatctc gctcccttct tcaccttcaa gtgttacggc 240
gtgagcccca ccaagctgaa tgacctctgc tttaccaatg tgtacgccga cagctttgtg 300
attagaggcg acgaagtgag acagatcgcc cctgggcaga ccggcaacat tgctgactat 360
aattacaagc tgcctgatga cttcaccggc tgtgtgattg cctggaacag caacaagctg 420
gattccaagg tgtccggcaa ttacaactac ctctaccggc tgttcagaaa gtccaacctg 480
aagcccttcg aaagagacat ctccaccgag atttaccaag ccggcaacaa gccctgcaac 540
ggcgtggccg gcttcaactg ctattttccc ctgcgcagct atagcttcag acccacctac 600
ggcgtgggcc atcaacccta tagagtggtc gtgctgagct tcgagctgct gcacgccccc 660
gccaccgtct gtggccccaa gaagagcacc aacctggtca agaacaagag agtgcagccc 720
accgagagca tcgtgcggtt ccccaatatc accaacctct gccctttcga cgaggtgttc 780
aacgctacaa gattcgcttc cgtgtacgcc tggaacagaa aacggatttc caactgcgtg 840
gccgactaca gcgtgctcta caacctggcc cccttcttta cattcaaatg ctacggcgtg 900
agccccacaa agctgaatga cctgtgcttc accaacgtgt atgccgacag cttcgtgatc 960
agaggcgacg aggtcagaca gatcgctccc gggcagaccg ggaacatcgc cgactacaac 1020
tataagctgc ccgacgattt taccggctgc gtgatcgcct ggaatagcaa caagctcgac 1080
agcaaggtga gcggcaacta taattatctg taccggctgt ttcggaagag caacctgaag 1140
ccctttgaga gagacatcag cacagagatc taccaagccg gcaacaaacc ctgtaacggc 1200
gtcgccggct ttaactgtta cttccccctg cgcagctatt ccttcagacc tacctatggc 1260
gtgggccatc agccctatag agtggtcgtg ctctccttcg aactgctgca cgctcctgcc 1320
accgtctgcg gccccaaaaa gtccacaaac ctggtgaaaa acaagcatca ccatcaccat 1380
cactga 1386
<210> 13
<211> 461
<212> PRT
<213> Artificial Sequence
<220>
<223> 合成多肽
<220>
<221> MISC_FEATURE
<222> (1)..(461)
<223> Delta-Omicron嵌合RBD-dimer构建体的氨基酸序列
<400> 13
Met Ile His Ser Val Phe Leu Leu Met Phe Leu Leu Thr Pro Thr Glu
1 5 10 15
Ser Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
20 25 30
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
35 40 45
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
50 55 60
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
65 70 75 80
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
85 90 95
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
100 105 110
Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
115 120 125
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
130 135 140
Gly Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu
145 150 155 160
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
165 170 175
Lys Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln
180 185 190
Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg
195 200 205
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
210 215 220
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
225 230 235 240
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
245 250 255
Asp Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
260 265 270
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
275 280 285
Leu Ala Pro Phe Phe Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
290 295 300
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
305 310 315 320
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile
325 330 335
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
340 345 350
Ala Trp Asn Ser Asn Lys Leu Asp Ser Lys Val Ser Gly Asn Tyr Asn
355 360 365
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
370 375 380
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Asn Lys Pro Cys Asn Gly
385 390 395 400
Val Ala Gly Phe Asn Cys Tyr Phe Pro Leu Arg Ser Tyr Ser Phe Arg
405 410 415
Pro Thr Tyr Gly Val Gly His Gln Pro Tyr Arg Val Val Val Leu Ser
420 425 430
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
435 440 445
Thr Asn Leu Val Lys Asn Lys His His His His His His
450 455 460
<210> 14
<211> 1386
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的多核苷酸
<220>
<221> misc_feature
<222> (1)..(1386)
<223> Delta-Omicron嵌合RBD-dimer构建体的核苷酸序列
<400> 14
atgatccaca gcgtgttcct gctgatgttc ctcctgaccc ctaccgagag cagagtgcag 60
cccacagagt ccatcgtgcg gtttcccaat attacaaacc tgtgtccctt cggggaagtg 120
ttcaacgcca caagattcgc tagcgtgtat gcttggaata gaaaaagaat ttccaactgc 180
gtggctgact acagcgtgct gtacaactcc gctagcttca gcaccttcaa gtgctatggg 240
gtgagcccca ccaagctgaa cgatctgtgc tttaccaatg tgtatgccga cagcttcgtg 300
attagagggg atgaggtccg gcagatcgct cccggccaaa ccggcaagat cgccgactac 360
aattacaagc tccccgatga ctttaccggc tgcgtgatcg cctggaatag caacaacctg 420
gactccaagg tgggcggcaa ttacaattac agatacagac tgtttcggaa gtccaacctg 480
aaacccttcg agcgggatat ctccaccgaa atctaccaag ccgggagcaa gccctgtaac 540
ggcgtggagg gcttcaactg ctactttccc ctgcagagct acggctttca gcccaccaac 600
ggcgtgggct atcagcccta cagagtggtc gtgctcagct tcgagctgct ccatgctccc 660
gctaccgtgt gcggccccaa gaaatccacc aacctggtga agaacaagcg ggtgcagccc 720
accgagagca tcgtcagatt ccccaacatc accaacctgt gccccttcga cgaggtgttc 780
aacgctacaa gattcgctag cgtgtacgcc tggaatagaa aaagaattag caattgtgtg 840
gctgattata gcgtcctgta caacctggcc cccttcttca cattcaagtg ctacggcgtc 900
tcccccacaa agctgaacga cctgtgcttc accaacgtgt acgccgatag ctttgtgatc 960
agaggcgacg aagtgagaca aatcgccccc gggcagaccg gcaacattgc cgactacaac 1020
tacaagctgc ccgacgactt caccggctgc gtcatcgctt ggaacagcaa taagctggac 1080
agcaaggtgt ccggcaacta taactacctc tacagactgt tcagaaagag caacctgaag 1140
cctttcgagc gggacatcag caccgagatt taccaagccg gcaacaagcc ctgcaatggc 1200
gtggctggct tcaactgtta cttccccctg cgcagctaca gcttcagacc cacctacggc 1260
gtgggccatc agccctaccg ggtggtcgtg ctgtccttcg agctgctgca cgctcccgcc 1320
accgtgtgtg gccccaagaa aagcaccaat ctggtcaaga acaagcatca ccatcaccat 1380
cactga 1386
<210> 15
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的寡核苷酸
<220>
<221> misc_feature
<222> (1)..(20)
<223> 用于检测SARS-CoV-2 Delta gRNA的上游引物
<400> 15
gaccccaaaa tcagcgaaat 20
<210> 16
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的寡核苷酸
<220>
<221> misc_feature
<222> (1)..(24)
<223> 用于检测SARS-CoV-2 Delta gRNA的下游引物
<400> 16
tctggttact gccagttgaa tctg 24
<210> 17
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的寡核苷酸
<220>
<221> misc_feature
<222> (1)..(24)
<223> 检测SARS-CoV-2 Delta gRNA的探针
<400> 17
accccgcatt acgtttggtg gacc 24
<210> 18
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的寡核苷酸
<220>
<221> misc_feature
<222> (1)..(24)
<223> 检测SARS-CoV-2 Omicron gRNA的探针
<400> 18
actccgcatt acgtttggtg gacc 24
<210> 19
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的寡核苷酸
<220>
<221> misc_feature
<222> (1)..(23)
<223> 用于检测SARS-CoV-2 Delta和Omicron sgRNA的上游引物
<400> 19
cgatctcttg tagatctgtt ctc 23
<210> 20
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的寡核苷酸
<220>
<221> misc_feature
<222> (1)..(22)
<223> 用于检测SARS-CoV-2 Delta和Omicron sgRNA的下游引物
<400> 20
atattgcagc agtacgcaca ca 22
<210> 21
<211> 26
<212> DNA
<213> Artificial Sequence
<220>
<223> 合成的寡核苷酸
<220>
<221> misc_feature
<222> (1)..(26)
<223> 用于检测SARS-CoV-2 Delta和Omicron sgRNA的探针
<400> 21
acactagcca tccttactgc gcttcg 26
Claims (25)
1.一种新型冠状病毒Delta和Omicron变异株的重组嵌合抗原，其特征在于：所述重组嵌合抗原的氨基酸序列包括：按照(A-B)-(A-B’)样式排列的氨基酸序列，其中：
A-B表示如SEQ ID NO:1所示的氨基酸序列，
A-B’表示如SEQ ID NO:2所示的氨基酸序列。
2.根据权利要求1所述的重组嵌合抗原，其特征在于：所述重组嵌合抗原的氨基酸序列如SEQ ID NO:3所示。
3.一种如权利要求1所述重组嵌合抗原的制备方法，其包括以下步骤：在编码如权利要求1所述重组嵌合抗原的核苷酸序列的5’端加上编码信号肽的核苷酸序列，3’端加上编码组氨酸的核苷酸序列和终止密码子，进行克隆表达，筛选正确的重组子，然后将其转染表达系统的细胞进行表达，收集细胞培养上清，从中分离获得所述重组嵌合抗原。
4.根据权利要求3所述的制备方法，其特征在于：所述表达系统的细胞包括哺乳动物细胞、昆虫细胞、酵母细胞或细菌细胞。
5.根据权利要求4所述的制备方法，其特征在于：所述哺乳动物细胞包括HEK293T细胞、HEK293F细胞、Expi293F细胞或CHO细胞。
6.根据权利要求4所述的制备方法，其特征在于：所述细菌细胞包括大肠杆菌细胞。
7.一种多核苷酸，其编码如权利要求1-2任一项所述的重组嵌合抗原。
8.根据权利要求7所述的多核苷酸，其特征在于：所述多核苷酸为DNA或mRNA。
9.根据权利要求7所述的多核苷酸，其特征在于：所述多核苷酸为如SEQ ID NO:4所示的核苷酸序列。
10.一种核酸构建体，其包含如权利要求7-9任一项所述的多核苷酸，以及任选地，与所述多核苷酸可操作地连接的至少一个表达调控元件。
11.一种表达载体，其包含如权利要求10所述的核酸构建体。
12.一种转化的细胞，其包括如权利要求7-9任一项所述的多核苷酸、如权利要求10所述的核酸构建体或如权利要求11所述的表达载体。
13.如权利要求1-2任一项所述的重组嵌合抗原、如权利要求7-9任一项所述的多核苷酸、如权利要求10所述的核酸构建体、如权利要求11所述的表达载体或如权利要求12所述的转化的细胞在制备新型冠状病毒疫苗中的应用。
14.一种疫苗或免疫原性组合物，其包含如权利要求1-2任一项所述的重组嵌合抗原、如权利要求7-9任一项所述的多核苷酸、如权利要求10所述的核酸构建体、如权利要求11所述的表达载体或如权利要求12所述的转化的细胞，以及生理学可接受的媒介物、佐剂、赋形剂、载体和/或稀释剂。
15.根据权利要求14所述的疫苗或免疫原性组合物，其为新型冠状病毒重组蛋白疫苗，其包括如权利要求1-2任一项所述的重组嵌合抗原和佐剂。
16.根据权利要求15所述的疫苗或免疫原性组合物，其特征在于：所述佐剂为选自以下佐剂中的一种或多种：铝佐剂、MF59佐剂和类MF59佐剂。
17.根据权利要求14所述的疫苗或免疫原性组合物，其为新型冠状病毒DNA疫苗，其包括：
（1）真核表达载体；和
（2）构建入所述真核表达载体中的、编码如权利要求1-2任一项所述的重组嵌合抗原的DNA序列。
18.根据权利要求17所述的疫苗或免疫原性组合物，其特征在于：所述真核表达载体选自pGX0001、pVAX1、pCAGGS和pCDNA系列载体。
19.根据权利要求14所述的疫苗或免疫原性组合物，其为新型冠状病毒mRNA疫苗，其包括编码如权利要求1-2任一项所述的重组嵌合抗原的mRNA序列和脂质纳米颗粒。
20.根据权利要求14所述的疫苗或免疫原性组合物，其为新型冠状病毒-病毒载体疫苗，其包括：
（1）病毒骨架载体；和
（2）构建入所述病毒骨架载体中的、编码如权利要求1-2任一项所述的重组嵌合抗原的核酸序列。
21.根据权利要求20所述的疫苗或免疫原性组合物，其特征在于：所述病毒骨架载体选自以下病毒载体中的一种或几种：腺病毒载体、痘病毒载体、流感病毒载体、腺相关病毒载体。
22.根据权利要求14-21任一项所述的疫苗或免疫原性组合物，其特征在于，所述疫苗或免疫原性组合物为鼻喷剂、口服制剂、栓剂或胃肠外制剂的形式。
23.根据权利要求22所述的疫苗或免疫原性组合物，其特征在于：所述鼻喷剂选自气雾剂、喷雾剂和粉雾剂。
24.根据权利要求22所述的疫苗或免疫原性组合物，其特征在于：所述口服制剂选自片剂、粉末剂、丸剂、散剂、颗粒剂、细粒剂、软/硬胶囊剂、薄膜包衣剂、小丸剂、舌下片和膏剂。
25.根据权利要求22所述的疫苗或免疫原性组合物，其特征在于：所述胃肠外制剂为经皮剂、软膏剂、硬膏剂、外用液剂、可注射或可推注制剂。
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113999321A (zh) * | 2021-12-24 | 2022-02-01 | 中国科学院微生物研究所 | 一种具有广谱中和活性的新冠病毒抗体及其制备方法与应用 |
| CN114044821A (zh) * | 2022-01-10 | 2022-02-15 | 中国人民解放军军事科学院军事医学研究院 | 一种抗新冠病毒全人源广谱中和抗体zwc12及应用 |
| CN114150005A (zh) * | 2022-02-09 | 2022-03-08 | 广州恩宝生物医药科技有限公司 | 用于预防SARS-CoV-2奥密克戎株的腺病毒载体疫苗 |
| CN114181962A (zh) * | 2022-01-17 | 2022-03-15 | 北京翊博普惠生物科技发展有限公司 | 一种新型冠状病毒mRNA疫苗及其制备方法与应用 |
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| WO2021239147A1 (zh) * | 2020-05-29 | 2021-12-02 | 中国科学院微生物研究所 | 一种β冠状病毒抗原、β冠状病毒二联疫苗及其制备方法和应用 |
| CN114057894B (zh) * | 2021-06-18 | 2022-08-05 | 国药中生生物技术研究院有限公司 | 一种产生广谱交叉中和活性的重组新型冠状病毒rbd三聚体蛋白疫苗、其制备方法和应用 |
| CN114634556B (zh) * | 2022-04-01 | 2023-12-19 | 中国科学院微生物研究所 | 一种新冠病毒Delta和Omicron变异株嵌合抗原、其制备方法和应用 |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113999321A (zh) * | 2021-12-24 | 2022-02-01 | 中国科学院微生物研究所 | 一种具有广谱中和活性的新冠病毒抗体及其制备方法与应用 |
| CN114044821A (zh) * | 2022-01-10 | 2022-02-15 | 中国人民解放军军事科学院军事医学研究院 | 一种抗新冠病毒全人源广谱中和抗体zwc12及应用 |
| CN114181962A (zh) * | 2022-01-17 | 2022-03-15 | 北京翊博普惠生物科技发展有限公司 | 一种新型冠状病毒mRNA疫苗及其制备方法与应用 |
| CN114150005A (zh) * | 2022-02-09 | 2022-03-08 | 广州恩宝生物医药科技有限公司 | 用于预防SARS-CoV-2奥密克戎株的腺病毒载体疫苗 |
Non-Patent Citations (1)
| Title |
|---|
| 新宇等.新型冠状病毒变异株以及疫苗研究现况.安徽预防医学杂志.2022,第28卷(第1期),参见第1-12页. * |
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