SEC-MALS验证的新冠突变株三聚体蛋白

SARS-CoV-2 Spike蛋白承担着病毒与宿主细胞膜受体结合及膜融合功能,是宿主中和抗体的重要作用位点以及疫苗设计的关键靶点。Spike蛋白含有2个不同功能的亚基:S1亚基通常呈球形,含有受体结合结构域,可与宿主受体结合;S2亚基促进病毒与宿主细胞的膜融合过程。

Spike蛋白通常以三聚体的形式存在,与宿主受体结合的过程会导致三聚体蛋白的构象变化。未结合宿主受体时,spike三聚体以亚稳定融合前构象(Prefusion)存在;结合ACE2后,S1亚基脱落,由S2亚基形成融合后构象(Postfusion)。2020年2月15日,美国NIH和德州大学的研究人员通过冷冻电镜技术首次解析新冠病毒S蛋白三聚体结构,发现S蛋白可利用构象变化来隐藏或暴露与ACE2结合的位点。随后,华盛顿大学的David Veesler研究团队发现S蛋白三聚体顶端存在多个不同的构象。研究人员利用Cryo-EM方法研究了S三聚体在病毒颗粒表面上的结构、构象和分布。约97%的S三聚体是融合前构象,仅仅3%是融合后构象。因为融合后的构象缺失了包含最重要的受体结合域的S1亚基,且有研究表明Spike三聚体蛋白的构象极不稳定,因此获得稳定的融合前构象三聚体S蛋白对于新冠病毒的科学研究、疫苗开发、中和抗体检测和诊断试剂开发至关重要。

义翘神州已成功开发出高稳定性的三聚体S蛋白,涵盖了多种重要的新冠突变体,如Alpha(B.1.1.7)、Beta(B.1.351)、Delta(B.1.617.2)、Kappa(B.1.617.1)等。三聚体S蛋白的纯度、聚体形式和活性分别经过SDS-PAGE、SEC-MALS和ELISA进行验证。且经过若干次冻融和多批次测定对稳定性进行检测,结果表明已开发的三聚体S蛋白均具有高稳定。

  • SARS-CoV-2 Spike (D614G) Trimer (ECD, His tag) (Cat: 40589-V08H8)

    SEC-MALS Validated

    The purity of SARS-CoV-2 Spike Trimer,His Tag (Cat. 40589-V08H8) was more than 90% and the molecular weight of this protein is around 400-540kDa verified by SEC-MALS.

    ACE2 Binding (ELISA)
     
    SDS-PAGE

    Immobilized human ACE2 protein (mFc tag) (Cat:10108-H05H) at 2 μg/mL (100 μL/well) can bind spike trimer (Cat:40589-V08H8), the EC50 is 4-24 ng/mL.

    Purity: > 90 %

  • SARS-CoV-2 B.1.617.2 Spike Trimer (ECD, His tag) (Cat: 40589-V08H10)

    SEC-MALS Validated

    The purity of SARS-CoV-2 Spike Trimer, His Tag (Cat. 40589-V08H10) was more than 90% and the molecular weight of this protein is around 420-520kDa verified by SEC-MALS.

    ACE2 Binding (ELISA)
     
    SDS-PAGE

    Immobilized human ACE2 protein (mFc tag) (Cat:10108-H05H) at 2 μg/mL (100 μL/well) can bind spike trimer (Cat:40589-V08H10), the EC50 is 4-24 ng/mL.

    Purity: > 90 %

新冠突变体Spike三聚体蛋白列表

货号 产品描述 突变位点
40589-V08H10 SARS-CoV-2 B.1.617.2 Spike S1+S2 trimer Protein (ECD, His tag) T19R, 156-157 deletion, R158D, L452R, T478K, D614G, F817P, A892P, A899P, A942P, D950N, K986P, V987P and furin cleavage site mutants
40589-V08H11 SARS-CoV-2 B.1.617.1 Spike S1+S2 trimer Protein (ECD, His tag) G142D, E154K, L452R, E484Q, D614G, F817P, A892P, A899P, A942P, K986P, V987P, Q1070H, H1101D and furin cleavage site mutants
40589-V08H12 SARS-CoV-2 B.1.1.7 Spike S1+S2 trimer Protein (ECD, His tag) (69-70, 145 deletion), N501Y, A570D, D614G, T716I, F817P, A892P, A899P, A942P, S982A, K986P, V987P, D1118H and furin cleavage site mutants
40589-V08H13 SARS-CoV-2 B.1.351 Spike S1+S2 trimer Protein (ECD, His tag) L18F, D80A, D215G, 241-243 deletion, R246I, K417N, E484K, N501Y, D614G, A701V, F817P, A892P, A899P, A942P, K986P, V987P and furin cleavage site mutants
40589-V08H8 SARS-CoV-2 Spike S1+S2 (D614G) trimer Protein (ECD, His tag) D614G, F817P, A892P, A899P, A942P, K986P, V987P and furin cleavage site mutants
40589-V08H9 SARS-CoV-2 B.1.618 Spike S1+S2 trimer Protein (ECD, His tag) H49Y, 145-146 deletion, E484K, D614G, F817P, A892P, A899P, A942P, K986P, V987P and furin cleavage site mutants