CN112538111B - 新冠病毒单链抗体及质控品和制备方法 - Google Patents
新冠病毒单链抗体及质控品和制备方法 Download PDFInfo
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Abstract
本发明涉及一种新冠病毒单链抗体及质控品和制备方法,从N段到C端依次包括轻链可变区和重链可变区,所述轻链可变区的氨基酸序列如SEQ ID NO:1所示,所述重链可变区的氨基酸序列如SEQ ID NO:2所示。本发明的新冠病毒单链抗体灵敏度高、稳定性好,非常适合质控品的制备。而且,在此可变区序列基础上进一步使用基因重组技术构建形成无CH1片段的人鼠嵌合IgM抗体,从而提高分子稳定性和特异性结合能力。本发明制备的新冠病毒单链抗体可应用于化学发光平台进行质控品检测,可以有效测定活性,能克服一般的2019‑nCoV IgM抗体检测灵敏度差、特异性低的缺陷。
Description
技术领域
本发明涉及免疫检测技术领域,特别是涉及一种新冠病毒单链抗体及质控品和制备方法。
背景技术
2019新型冠状病毒(2019-nCoV),2020年1月12日被世界卫生组织命名为“2019-nCoV”,2020年2月11日被国际病毒分类委员会命名为“严重急性呼吸综合征冠状病毒2(SARS-CoV-2)”。新型冠状病毒是以前从未在人体中发现的冠状病毒新毒株113。冠状病毒是一个大型病毒家族,已知可引起感冒以及中东呼吸综合征(MERS)和严重急性呼吸综合征(SARS)等较严重疾病。2019-nCoV与SARS病毒在整个基因组的相似度约为80%。从目前的认知来看,在毒性上,2019-nCoV比SARS弱,而在传染性上,因2019-nCoV潜伏期长,且无症状感染者也存在传染性,且人群全部易感,导致传播控制难度更大。基于日前的流行病学调查,该病潜伏期一般为3~7天,最短潜伏期是1天,最长潜伏期为14天,在潜伏期间依然具有传染性。该病能够在人与人之间传播,主要通过飞沫和接触传播,在密闭不通风的场所可能存在气溶胶传播风险。免疫能力低下的人群感染后病情较严重,儿童及婴幼儿也有发病。人感染了2019-nCoV后常见体征有呼吸道症状、发热、咳嗽、气促和呼吸困难等,在较严重病例中,感染可导致肺炎、严重急性呼吸综合征、肾衰竭,甚至死亡。
感染2019-nCoV后,人体在约7天左右会产生针对病毒的IgM抗体,约14天左右会产生针对病毒的IgG抗体。产生的抗体是一种保护性的抗体,能促进人体从病毒感染中恢复和自愈。2019-nCoV抗体检测已广泛应用于ELISA、胶体金、侧向层析、化学发光等平台,但一般的质控抗体因稳定性和灵敏度不足等原因,无法进行准确有效的测定。
发明内容
基于此,有必要提供一种稳定性和灵敏度较好的新冠病毒单链抗体。
一种新冠病毒单链抗体,从N端到C端依次包括轻链可变区和重链可变区,所述轻链可变区的氨基酸序列如SEQ ID NO:1所示,所述重链可变区的氨基酸序列如SEQ ID NO:2所示。
在其中一个实施例中,还包括依次连接于所述重链可变区的C端的人IgM抗体轻链恒定区和人IgM抗体重链恒定区。
在其中一个实施例中,所述人IgM抗体轻链恒定区的氨基酸序列如SEQ ID NO:3所示,所述人IgM抗体重链恒定区的氨基酸序列如SEQ ID NO:4所示。
本发明还提供了一种分离的核酸,其编码上述新冠病毒单链抗体。
在其中一个实施例中,包括序列如SEQ ID NO:5和SEQ ID NO:6所示的核苷酸片段。
在其中一个实施例中,包括序列如SEQ ID NO:7和SEQ ID NO:8所示的核苷酸片段。
本发明还提供了一种新冠病毒抗体质控品的制备方法,包括以下步骤:
将新冠病毒抗体与冻干保护溶液混合,得到抗体溶液;
将所述抗体溶液依次进行预冻处理、一次干燥处理和二次干燥处理,得到所述新冠病毒抗体质控品;
所述预冻处理的条件参数依次为:隔板温度-5℃~-15℃处理35~45min,隔板温度-45℃~-55℃处理80~100min,隔板温度-20℃~-30℃处理80~100min,隔板温度-50℃~-60℃处理80~100min;所述一次干燥处理的条件参数依次为:隔板温度-50℃~-60℃、真空度0.2mbar~0.4mbar条件下处理6~10s,隔板温度-50℃~-60℃、真空度0.08mbar~0.12mbar条件下处理1~3s,隔板温度-25℃~-35℃、真空度0.08mbar~0.12mbar条件下处理20~21小时,隔板温度-5℃~-15℃、真空度0.08mbar~0.12mbar条件下处理25min~35min,隔板温度-5℃~-15℃、真空度0.06mbar~0.10mbar条件下处理50~70min;所述二次干燥处理的条件参数为:隔板温度5℃~15℃、真空度0mbar~0.005mbar条件下处理5~6小时。
在其中一个实施例中,所述冻干保护溶液含有以下浓度的组分:0.02M~0.06MTris-HCl、0.006M~0.010M Tris、0.6wt%~1.2wt%氯化钠、2wt%~6wt%牛血清白蛋白、6wt%~10wt%甘露醇、1wt%~3wt%海藻糖。
在其中一个实施例中,所述冻干保护溶液还含有以下浓度的组分:0.8wt%~1.2wt%明胶和0.8wt%~1.2wt%羟乙基淀粉。
本发明还提供了一种新冠病毒抗体质控品,其根据上述制备方法制备得到。
本发明的新冠病毒单链抗体灵敏度高、稳定性好,非常适合质控品的制备。而且,在此可变区序列基础上进一步使用基因重组技术构建形成无CH1片段的人鼠嵌合IgM抗体,从而提高分子稳定性和特异性结合能力。本发明制备的新冠病毒单链抗体可应用于化学发光平台进行质控品检测,可以有效测定活性,能克服一般的2019-nCoV IgM抗体检测灵敏度差、特异性低的缺陷。
本发明采用自主研究的新型冻干程序,运用分步升降隔板温度和分步改变真空度相结合的技术改进传统的快速冻干程序,结合质控抗体加工制备可得到临床免疫冻干粉形态的新冠病毒抗体质控品,在体外存放稳定性好,具有保存周期长、黏稠度低、容易复溶与混匀、基质效应较小等优势,对检测系统和试剂盒测试结果的精密度非常重要。该质控品适配于化学发光检测平台,可与对应的检测系统和试剂盒配套使用。本发明还添加了明胶和羟乙基淀粉双重高分子聚合物作为冻干保护剂,从而提高玻璃化转变温度,可进一步改善新冠病毒抗体质控品的冻干形态。
具体实施方式
为了便于理解本发明,下面将对本发明进行更全面的描述,并给出了本发明的较佳实施例。但是,本发明可以以许多不同的形式来实现,并不限于本文所描述的实施例。相反地,提供这些实施例的目的是使对本发明的公开内容的理解更加透彻全面。
除非另有定义,本文所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本文中在本发明的说明书中所使用的术语只是为了描述具体的实施例的目的,不是旨在于限制本发明。本文所使用的术语“和/或”包括一个或多个相关的所列项目的任意的和所有的组合。
本发明一实施例的新冠病毒单链抗体,从N端到C端依次包括轻链可变区和重链可变区,轻链可变区的氨基酸序列如SEQ ID NO:1所示,重链可变区的氨基酸序列如SEQ IDNO:2所示。
在一个具体示例中,新冠病毒单链抗体还包括依次连接于上述重链可变区的C端的人IgM抗体轻链恒定区和人IgM抗体重链恒定区。2019-nCoV IgM抗体是体液免疫中最早出现的特异性抗体,当病毒感染人体时,IgM抗体检测对病毒感染的早期诊断具有重要意义。
在一个具体示例中,上述人IgM抗体轻链恒定区的氨基酸序列如SEQ ID NO:3所示,上述人IgM抗体重链恒定区的氨基酸序列如SEQ ID NO:4所示。如此,使用基因重组技术构建形成无CH1片段的人鼠嵌合IgM抗体,利用此手段可提高分子稳定性和特异性结合能力。
本发明的新冠病毒单链抗体灵敏度高、稳定性好,非常适合质控品的制备。而且,在此可变区序列基础上进一步使用基因重组技术构建形成无CH1片段的人鼠嵌合IgM抗体,从而提高分子稳定性和特异性结合能力。本发明制备的新冠病毒单链抗体可应用于化学发光平台进行质控品检测,可以有效测定活性,能克服一般的2019-nCoV IgM抗体检测灵敏度差、特异性低的缺陷。
本发明一实施例的分离的核酸,其编码上述新冠病毒单链抗体。
在一个具体示例中,上述分离的核酸包括序列如SEQ ID NO:5和SEQ ID NO:6所示的核苷酸片段。可以理解,由于密码子的简并性,能够表达同一蛋白的核酸序列具有多种形式,以上为经过密码子优化的核酸序列,但不限于此。
在一个具体示例中,上述分离的核酸包括序列如SEQ ID NO:7和SEQ ID NO:8所示的核苷酸片段,分别编码上述人IgM抗体轻链恒定区和人IgM抗体重链恒定区。
本发明一实施例的重组表达载体,该重组表达载体用于表达上述新冠病毒单链抗体。
可以理解,重组表达载体中还可包含基因工程中常用的调控元件,例如增强子、启动子等其他表达控制元件(例如转录终止信号、或者多腺苷酸化信号和多聚U序列等)。在一个具体示例中,初始载体可以选择pCMVp-NEO-BAN载体。
本发明一实施例的宿主细胞,其含有上述重组表达载体。在一个具体示例中,宿主细胞为HEK293细胞。
本发明一实施例的新冠病毒抗体质控品的制备方法,包括以下步骤:
将新冠病毒抗体与冻干保护溶液混合,得到抗体溶液。
将抗体溶液依次进行预冻处理、一次干燥处理和二次干燥处理,得到新冠病毒抗体质控品。
上述预冻处理的条件参数依次为:隔板温度-5℃~-15℃处理35~45min,隔板温度-45℃~-55℃处理80~100min,隔板温度-20℃~-30℃处理80~100min,隔板温度-50℃~-60℃处理80~100min。上述一次干燥处理的条件参数依次为:隔板温度-50℃~-60℃、真空度0.2mbar~0.4mbar条件下处理6~10s,隔板温度-50℃~-60℃、真空度0.08mbar~0.12mbar条件下处理1~3s,隔板温度-25℃~-35℃、真空度0.08mbar~0.12mbar条件下处理20~21小时,隔板温度-5℃~-15℃、真空度0.08mbar~0.12mbar条件下处理25min~35min,隔板温度-5℃~-15℃、真空度0.06mbar~0.10mbar条件下处理50~70min。上述二次干燥处理的条件参数为:隔板温度5℃~15℃、真空度0mbar~0.005mbar条件下处理5~6小时。
本发明采用自主研究的新型冻干程序,运用分步升降隔板温度和分步改变真空度相结合的技术改进传统的快速冻干程序,结合质控抗体加工制备可得到临床免疫冻干粉形态的新冠病毒抗体质控品,在体外存放稳定性好,具有保存周期长、黏稠度低、容易复溶与混匀、基质效应较小等优势,对检测系统和试剂盒测试结果的精密度非常重要。该质控品适配于化学发光检测平台,可与对应的检测系统和试剂盒配套使用。
在一个具体示例中,上述抗体溶液中新冠病毒抗体的浓度为5AU/mL~24AU/mL。
在一个具体示例中,冻干保护溶液含有以下浓度的组分:0.02M~0.06MTris-HCl、0.006M~0.010M Tris、0.6wt%~1.2wt%氯化钠、2wt%~6wt%牛血清白蛋白、6wt%~10wt%甘露醇、1wt%~3wt%海藻糖。
在一个具体示例中,冻干保护溶液还含有以下浓度的组分:0.8wt%~1.2wt%明胶和0.8wt%~1.2wt%羟乙基淀粉。如此,添加了明胶和羟乙基淀粉双重高分子聚合物作为冻干保护剂,从而提高玻璃化转变温度,可进一步改善新冠病毒抗体质控品的冻干形态。
以下为具体实施例。
实施例1
1.全鼠源IgM单克隆抗体制备
动物免疫:根据中国CDC公布的N基因序列进行蛋白合成,通过安徽通用生物合成N蛋白,N基因序列如下所示。以制备的2019-nCoV N蛋白作为免疫原免疫小鼠,制备全鼠源IgM单克隆抗体。将N蛋白与弗氏佐剂等量混合充分乳化后注射3只Balb/c小鼠,每只小鼠25μg,共免疫3次,间隔时间15天。
N基因序列:
GGGGAACTTCTCCTGCTAGAATGGCTGGCAATGGCGGTGATGCTGCTCTTGCTTTGCTGCTGCTTGACAGATTGAACCAGCTTGAGAGCAAAATGTCTGG
细胞融合:融合前3天,小鼠腹腔注射偶联蛋白25μg,取出免疫小鼠的脾脏细胞,将2.0×107个SP2/0骨髓瘤细胞与2.0×108个经免疫的Balb/c小鼠的脾细胞混匀,离心,弃上清,轻微振荡混匀,置于37℃水浴锅中,在90秒内滴加1mL体积浓度为50%的PEG-1500水溶液,然后滴加20mL的1640培养基,离心,弃上清,再洗一次,离心,弃上清,得到杂交瘤细胞。使用HAT选择培养基在96孔细胞培养板上选择杂交瘤细胞,镜下检测总融合率>95%。包被N蛋白对单克隆细胞孔的上清进行检测,挑取OD450>0.8的孔内细胞进行亚克隆,最终获得对天然N蛋白反应阳性率>99%的细胞克隆,作为分泌抗人N蛋白的IgM单克隆抗体的杂交瘤细胞株。
细胞克隆:对获得的阳性细胞株进行克隆,使用有限稀释法,克隆3次,最后得到产生高效价的抗人N蛋白的IgM单克隆抗体杂交瘤细胞系5株,扩大培养,冻存。
腹水制备:取6~8周龄的雌性Balb/c小鼠用石蜡处理10天后,取杂交瘤细胞以2×106个细胞/只小鼠进行腹腔注射。7天后从小鼠腹腔中获取富含IgM抗体的腹水,测定腹水效价>105。
抗体纯化:采用protein G亲和层析法。首先制备protein G亲和层析柱,用PBS平衡柱子后,取腹水过柱,然后用PBS清洗柱子,以50nmol/L的甘氨酸盐酸盐溶液洗脱,收集洗脱液,测定各收集管的OD值,保留峰值区的洗脱液,洗脱液经透析后收集。经SDS-PAGE电泳鉴定为纯化的鼠源IgM单克隆抗体,纯度为98%以上。抗体的可变区序列如下表1所示。
表1
2.扩增合成鼠源IgM单克隆抗体的可变区基因和人源IgM抗体的恒定区基因
取鼠源IgM单克隆抗体基因作为扩增模板,利用一对轻链可变区引物和一对重链可变区引物,以四种核苷酸dNTPs作为原料,在DNA聚合酶作用下扩增合成鼠源可变区基因。
取人源IgM抗体基因作为扩增模板,利用一对轻链恒定区引物和一对重链恒定区引物,以四种核苷酸dNTPs作为原料,在DNA聚合酶作用下扩增合成人源恒定区基因。以人源恒定区基因为模板,扩增合成无CH1片段的恒定区基因,如下表2所示。
表2
3.构建无CH1片段的鼠-人IgM轻重链基因表达载体
通过基因重组,将人源的无CH1片段的恒定区基因和鼠源的可变区基因进行拼接,拼接的抗体基因序列见表3,克隆到表达载体pCMVp-NEO-BAN,构建出无CH1片段的人-鼠IgM嵌合抗体基因表达载体。
表3
4.IgM嵌合抗体表达与纯化
IgM嵌合抗体表达:将无CH1片段的人-鼠IgM嵌合抗体基因表达载体质粒转染进293细胞,收集细胞上清,获得无CH1片段的人鼠嵌合的2019-nCoV IgM抗体。
IgM嵌合抗体纯化:采用protein G亲和层析法。首先制备protein G亲和层析柱,用PBS平衡柱子后,取腹水过柱,然后用PBS清洗柱子,以50nmol/L的甘氨酸盐酸盐溶液洗脱,收集洗脱液,测定各收集管的OD值,保留峰值区的洗脱液,洗脱液经透析后收集。经SDS-PAGE电泳鉴定为纯化的IgM嵌合抗体,纯度为98%以上。
5.IgM嵌合抗体活性检测
纯化的IgM嵌合抗体用Tris缓冲液分别稀释10倍、50倍和100倍,选用纯化的普通单克隆抗体和多克隆抗体作为对照,均用Tris缓冲液分别稀释10倍、50倍和100倍。选用公司生产的新型冠状病毒IgM抗体测定试剂盒,在化学发光平台检测抗体活性,可见IgM嵌合抗体活性较高,回算后的浓度高于IgM单克隆抗体和多克隆抗体,发光值和浓度值见表4。
表4
纯化的含可变区和不含CH1恒定区的IgM抗体用Tris缓冲液分别稀释10倍、50倍、100倍、300倍和600倍,选用纯化的仅含可变区的IgM抗体和纯化的含可变区和完整恒定区的IgM抗体作为对照,均用Tris缓冲液分别稀释10倍、50倍、100倍、300倍和600倍。选用公司生产的新型冠状病毒IgM抗体测定试剂盒,在化学发光平台检测抗体亲和力和灵敏度,可见含可变区和不含CH1恒定区的IgM抗体亲和力和灵敏度均高于仅含可变区的IgM抗体和含可变区和完整恒定区的IgM抗体,亲和力和灵敏度的对比结果见表5和表6。
表5
表6
以纯化的含可变区和不含CH1恒定区的IgM抗体原液作为对照组,选用纯化的仅含可变区的IgM抗体和纯化的含可变区和完整恒定区的IgM抗体作为对照,选用公司生产的抗RA33 IgG抗体,新型冠状病毒IgG抗体和新型冠状病毒IgM抗体测定试剂盒,在化学发光平台检测抗体特异性,特异性的对比结果见表7。
表7
实施例2制备新冠病毒IgM抗体质控品
原料处理和浓度确认:把上述IgM嵌合抗体原料从保存条件下冰箱取出解冻,在十万级生产车间的生物安全柜对原料进行处理,用乙型肝炎病毒表面抗原测定试剂盒、丙型肝炎病毒抗体测定试剂盒、人类免疫缺陷病毒抗原抗体测定试剂盒以及梅毒螺旋体抗体测定试剂盒检测,选择检测结果均呈阴性或无反应性的原料,再测定2019-nCoV IgM嵌合抗体原料的浓度。然后进行离心和过滤,得到澄清的抗体原料,并在十万级生产车间确认抗体浓度。
溶液配制:在十万级生产车间进行冻干溶液的配制。配制过程如下:以1L溶液为例,0.04M Tris-HCl+0.008M Tris+0.9%氯化钠+4%牛血清白蛋白+1%明胶+1%羟乙基淀粉+0.05%PC300+0.02%PS-3+8%甘露醇+2%海藻糖+3%F012,按照各个组分含量计算称取量。先加Tris-HCl、Tris、氯化钠、PC300等组分,加800mL纯水溶解后将溶液pH调整到7.4±0.5范围内,提前加热溶解明胶,然后加牛血清白蛋白和羟乙基淀粉微调溶液pH至7.4±0.5范围内,加纯水补齐至1L。
质控品配制、分装和冻干:若处理后的IgM嵌合抗体原料符合公司要求,则继续在十万级生产车间进行质控品的配制、分装和冻干。往溶液中投入2019-nCoV IgM嵌合抗体材料配制成浓度为(5±30%)AU/mL和(20±20%)AU/mL)的双浓度质控品,使用新型冻干程序和常规冻干程序进行冻干,程序见表8,冻干结束后真空下压盖。分别验证两种程序下冻干的IgM抗体质控品的稳定性性能,发现使用新型冻干程序冻干得到的质控品热稳定性和复溶稳定性均优于常规冻干程序,结果见表9和表10。
表8
表9
表10
质控品赋值:取样复溶并在普通车间进行赋值。赋值步骤如下:准备5台化学发光测定仪,在每台仪器上使用经产品校准后的新冠IgM抗体测定试剂盒(化学发光法)分别重复测定质控品共测5天,每天测试4次。5天测试完成后,每台仪器每个质控品浓度得出20个测试数据。检测结果依据格拉布斯(Grubbs)准则剔除异常值,剔除离群值的个数不得超过2.5%,当离群值超过2.5%时,应怀疑是否为方法不稳定或操作者不熟悉所致。此时,不可使用此次试验的数据,检查问题和解决问题后重新开始新的评估实验。将检测结果剔除异常值后计算剩下数据的均值X,标准差SD,确定质控品靶值范围(X±3SD)。
半成品检验和组装:赋值的下一步进行半成品检验,检验质控品的准确度和重复性,若检验通过则进行产品组装和成品检验,若不通过则重新进行质控品的赋值并进行半成品检或者将其作不合格品处理。
成品检验和入库:组装的下一步进行成品检验,检验质控品的准确度和重复性,检验通过后,则进行成品入库,否则当不合格品处理。
实施例3新冠病毒IgM抗体质控品的稳定性验证
为了验证2019-nCoV IgM质控品在高温等极端环境中的性能表现,本发明将产品放在37℃条件下进行热处理10天,2~8℃条件下保存的产品作为对照;同时,为了确定2019-nCoV IgM质控品复溶后在不同储存条件下的稳定性,本发明将产品复溶后分别置于20~25℃、2~8℃和-20℃保存3天、30天和90天,2~8℃条件下保存的产品作为对照。选用公司生产的新型冠状病毒IgM抗体测定试剂盒,在化学发光平台测定各个条件下2019-nCoVIgM质控品的发光值和浓度值,结果稳定性较好,相对偏差均在±10%以内,如表11和表12所示。
表11
表12
以上所述实施例的各技术特征可以进行任意的组合,为使描述简洁,未对上述实施例中的各个技术特征所有可能的组合都进行描述,然而,只要这些技术特征的组合不存在矛盾,都应当认为是本说明书记载的范围。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
序列表
<110> 深圳市亚辉龙生物科技股份有限公司
<120> 新冠病毒单链抗体及质控品和制备方法
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<400> 7
cgtacggtgg ctgcaccatc tgtcttcatc ttcccgccat ctgatgagca gttgaaatct 60
ggaactgcct ctgttgtgtg cctgctgaat aacttctatc ccagagaggc caaagtacag 120
tggaaggtgg ataacgccct ccaatcgggt aactcccagg agagtgtcac agagcaggac 180
agcaaggaca gcacctacag cctcagcagc accctgacgc tgagcaaagc agactacgag 240
aaacacaaag tctacgcctg cgaagtcacc catcagggcc tgagctcgcc cgtcacaaag 300
agcttcaaca ggggagagtg t 321
<210> 8
<211> 1047
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
gtgattgctg agctgcctcc caaagtgagc gtcttcgtcc caccccgcga cggcttcttc 60
ggcaaccccc gcaagtccaa gctcatctgc caggccacgg gtttcagtcc ccggcagatt 120
caggtgtcct ggctgcgcga ggggaagcag gtggggtctg gcgtcaccac ggaccaggtg 180
caggctgagg ccaaagagtc tgggcccacg acctacaagg tgaccagcac actgaccatc 240
aaagagagcg actggctcag ccagagcatg ttcacctgcc gcgtggatca caggggcctg 300
accttccagc agaatgcgtc ctccatgtgt gtccccgatc aagacacagc catccgggtc 360
ttcgccatcc ccccatcctt tgccagcatc ttcctcacca agtccaccaa gttgacctgc 420
ctggtcacag acctgaccac ctatgacagc gtgaccatct cctggacccg ccagaatggc 480
gaagctgtga aaacccacac caacatctcc gagagccacc ccaatgccac tttcagcgcc 540
gtgggtgagg ccagcatctg cgaggatgac tggaattccg gggagaggtt cacgtgcacc 600
gtgacccaca cagacctgcc ctcgccactg aagcagacca tctcccggcc caagggggtg 660
gccctgcaca ggcccgatgt ctacttgctg ccaccagccc gggagcagct gaacctgcgg 720
gagtcggcca ccatcacgtg cctggtgacg ggcttctctc ccgcggacgt cttcgtgcag 780
tggatgcaga gggggcagcc cttgtccccg gagaagtatg tgaccagcgc cccaatgcct 840
gagccccagg ccccaggccg gtacttcgcc cacagcatcc tgaccgtgtc cgaagaggaa 900
tggaacacgg gggagaccta cacctgcgtg gtggcccatg aggccctgcc caacagggtc 960
accgagagga ccgtggacaa gtccaccggt aaacccaccc tgtacaacgt gtccctggtc 1020
atgtccgaca cagctggcac ctgctac 1047
Claims (10)
1.一种新冠病毒单链抗体,其特征在于,从N端到C端依次包括轻链可变区和重链可变区,所述轻链可变区的氨基酸序列如SEQ ID NO:1所示,所述重链可变区的氨基酸序列如SEQ ID NO:2所示。
2.根据权利要求1所述的新冠病毒单链抗体,其特征在于,还包括依次连接于所述重链可变区的C端的人IgM抗体轻链恒定区和人IgM抗体重链恒定区。
3.根据权利要求2所述的新冠病毒单链抗体,其特征在于,所述人IgM抗体轻链恒定区的氨基酸序列如SEQ ID NO:3所示,所述人IgM抗体重链恒定区的氨基酸序列如SEQ ID NO:4所示。
4.一种分离的核酸,其特征在于,其编码权利要求1~3任一项所述的新冠病毒单链抗体。
5.根据权利要求4所述的分离的核酸,其特征在于,包括序列如SEQ ID NO:5和SEQ IDNO:6所示的核苷酸片段。
6.根据权利要求4所述的分离的核酸,其特征在于,包括序列如SEQ ID NO:7和SEQ IDNO:8所示的核苷酸片段。
7.一种新冠病毒抗体质控品的制备方法,其特征在于,包括以下步骤:
将新冠病毒抗体与冻干保护溶液混合,得到抗体溶液;所述新冠病毒抗体为权利要求1~3任一项所述的新冠病毒单链抗体;
将所述抗体溶液依次进行预冻处理、一次干燥处理和二次干燥处理,得到所述新冠病毒抗体质控品;
所述预冻处理的条件参数依次为:隔板温度-5℃~-15℃处理35~45min,隔板温度-45℃~-55℃处理80~100min,隔板温度-20℃~-30℃处理80~100min,隔板温度-50℃~-60℃处理80~100min;所述一次干燥处理的条件参数依次为:隔板温度-50℃~-60℃、真空度0.2mbar~0.4mbar条件下处理6~10s,隔板温度-50℃~-60℃、真空度0.08mbar~0.12mbar条件下处理1~3s,隔板温度-25℃~-35℃、真空度0.08mbar~0.12mbar条件下处理20~21小时,隔板温度-5℃~-15℃、真空度0.08mbar~0.12mbar条件下处理25min~35min,隔板温度-5℃~-15℃、真空度0.06mbar~0.10mbar条件下处理50~70min;所述二次干燥处理的条件参数为:隔板温度5℃~15℃、真空度0mbar~0.005mbar条件下处理5~6小时。
8.根据权利要求7所述的制备方法,其特征在于,所述冻干保护溶液含有以下浓度的组分:0.02M~0.06M Tris-HCl、0.006M~0.010M Tris、0.6wt%~1.2wt%氯化钠、2wt%~6wt%牛血清白蛋白、6wt%~10wt%甘露醇、1wt%~3wt%海藻糖。
9.根据权利要求8所述的制备方法,其特征在于,所述冻干保护溶液还含有以下浓度的组分:0.8wt%~1.2wt%明胶和0.8wt%~1.2wt%羟乙基淀粉。
10.一种新冠病毒抗体质控品,其特征在于,根据权利要求7~9任一项所述的制备方法制备得到。
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Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH03197867A (ja) * | 1989-12-26 | 1991-08-29 | Tokuyama Soda Co Ltd | 凝集反応用試薬 |
AU2945199A (en) * | 1998-03-18 | 1999-10-11 | Ronai, Peter | Amorphous glasses for stabilising sensitive products |
DE10361598A1 (de) * | 2003-12-24 | 2005-07-28 | Boehringer Ingelheim Pharma Gmbh & Co. Kg | Gefriergetrocknete Formulierung von Antikörperkonjugaten |
CN101644713A (zh) * | 2009-07-10 | 2010-02-10 | 杨维明 | 一种6-8万分子量范围的羟乙基淀粉的应用 |
CN101716343A (zh) * | 2008-10-09 | 2010-06-02 | 哈药集团生物工程有限公司 | 一种单克隆抗体的冻干制剂 |
CN101827608A (zh) * | 2007-06-14 | 2010-09-08 | 伊兰制药公司 | 冻干的免疫球蛋白制剂和制备方法 |
CN102813932A (zh) * | 2012-08-30 | 2012-12-12 | 青岛康地恩药业股份有限公司 | 鸭病毒性肝炎卵黄抗体冻干保护剂 |
CN111351318A (zh) * | 2020-03-07 | 2020-06-30 | 苏州德沃生物技术有限公司 | 抗体偶联乳胶的冻干方法 |
CN111620945A (zh) * | 2020-05-09 | 2020-09-04 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | 一种抗新型冠状病毒的单克隆抗体或其衍生体 |
-
2020
- 2020-12-09 CN CN202011430713.3A patent/CN112538111B/zh active Active
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH03197867A (ja) * | 1989-12-26 | 1991-08-29 | Tokuyama Soda Co Ltd | 凝集反応用試薬 |
AU2945199A (en) * | 1998-03-18 | 1999-10-11 | Ronai, Peter | Amorphous glasses for stabilising sensitive products |
DE10361598A1 (de) * | 2003-12-24 | 2005-07-28 | Boehringer Ingelheim Pharma Gmbh & Co. Kg | Gefriergetrocknete Formulierung von Antikörperkonjugaten |
CN101827608A (zh) * | 2007-06-14 | 2010-09-08 | 伊兰制药公司 | 冻干的免疫球蛋白制剂和制备方法 |
CN101716343A (zh) * | 2008-10-09 | 2010-06-02 | 哈药集团生物工程有限公司 | 一种单克隆抗体的冻干制剂 |
CN101644713A (zh) * | 2009-07-10 | 2010-02-10 | 杨维明 | 一种6-8万分子量范围的羟乙基淀粉的应用 |
CN102813932A (zh) * | 2012-08-30 | 2012-12-12 | 青岛康地恩药业股份有限公司 | 鸭病毒性肝炎卵黄抗体冻干保护剂 |
CN111351318A (zh) * | 2020-03-07 | 2020-06-30 | 苏州德沃生物技术有限公司 | 抗体偶联乳胶的冻干方法 |
CN111620945A (zh) * | 2020-05-09 | 2020-09-04 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | 一种抗新型冠状病毒的单克隆抗体或其衍生体 |
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