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新冠病毒相关蛋白和抑制剂筛选试剂盒

背景
2019年12月,湖北省武汉市首次集中发现不明原因肺炎,现已确诊为新型冠状病毒肺炎。冠状病毒是一类具有囊膜、基因组为线性单股正链RNA病毒,分为α、β、γ和δ四个属,此次新型冠状病毒属于β冠状病毒属(Betacoronavirus)。冠状病毒包含E (envelope protein)、M (membrane protein)、N (nucleocapsid protein)和S (spike protein)四种典型结构蛋白。其中,S蛋白与病毒的感染能力相关,是冠状病毒最重要的表面蛋白。S蛋白包含S1和S2两个功能性亚基,S1包含受体结合区域(RBD),负责识别和结合细胞受体,S2含有膜融合过程所需的基本元件,负责将S蛋白锚定在病毒膜上并介导病毒膜与宿主细胞膜的融合。所以S蛋白是介导冠状病毒进入宿主细胞的主要蛋白,也是感染后引起宿主免疫系统产生中和性抗体的主要靶标。
S蛋白在冠状病毒的表面会形成突出的同型三聚体,2020年2月份Science重磅报道了SARS-CoV-2 S蛋白三聚体的cyro-EM结构,不仅为解释SARS-CoV-2 S蛋白与ACE2蛋白的亲和力比SARS S高10-20倍提供了结构依据,也进一步阐释了S蛋白三聚体结构对于介导冠状病毒进入宿主细胞的重要作用。
蛋白的聚集状态和结构形式会影响其发挥正确的生物学功能,也会影响抗病毒药物的开发进展和治疗效果。为保证S蛋白的trimer结构正确,ACROBiosystems应用独特的技术开发了具有trimer结构的S蛋白,并使用SEC-MALS技术对蛋白的分子量和聚体形式进行了验证,蛋白产品经过SPR和BLI技术的动力学表征和亲和力的严格验证并确认其天然结合活性,免费提供Protocol。
目前市场尚无针对COVID-19的特定治疗方法,迫切需要开发针对COVID-19的疫苗、治疗性抗体或者小分子药物。ACRO基于已开发的生物素标记ACE2蛋白和新冠病毒蛋白,开发了SARS-CoV-2抑制剂筛选试剂盒,可用于SARS-CoV-2抑制剂的筛选,助力于药物研发。
蛋白分子量和活性验证数据
S三聚体蛋白的纯度高于85%,经MALS验证分子量为640-680 kDa
Fig.1 The purity of SARS-CoV-2 S protein (R683A, R685A), His Tag, active trimer (Cat. No. SPN-C52H8) was more than 85% and around 640-680 kDa verified by SEC-MALS.
SPR验证ACE2蛋白与SARS-CoV-2 S三聚体蛋白的亲和力常数为35.3 nM
Fig.2 Human ACE2, Fc Tag (Cat. No. AC2-H5257) Captured on CM5 chip via anti-human IgG Fc antibodies surface can bind SARS-CoV-2 S protein (R683A, R685A), His Tag, active trimer (Cat. No. SPN-C52H8) with an affinity constant of 35.3 nM as determined in a SPR assay (Biacore 8K).

Protocol

ELISA验证生物素化Human ACE2蛋白与SARS-CoV-2 S三聚体蛋白结合的线性范围为4-63 ng/mL
Fig.3 Immobilized SARS-CoV-2 S protein (R683A, R685A), His Tag, active trimer (Cat. No. SPN-C52H8) at 2 μg/mL (100 μL/well) can bind Biotinylated Human ACE2, His,Avitag (Cat. No. AC2-H82E6) with a linear range of 4-63 ng/mL.

Protocol

产品列表

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Kits  For research use only.
Molecule Cat. No. Species Product Description Protocol/Order
S1 protein TAS-K001 SARS-CoV-2 SARS-CoV-2 antibody titer assay kit (Spike protein S1)

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S protein RBD TAS-K002 SARS-CoV-2 SARS-CoV-2 antibody titer assay kit (Spike protein RBD)

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TAS-K003 SARS-CoV-2 Anti-SARS-CoV-2 neutralizing antibody titer serologic assay kit

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EP-105 SARS-CoV-2 SARS-CoV-2 Inhibitor Screening Kit

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S1 protein MBS-K001 SARS-CoV-2 SARS-CoV-2 Spike S1 protein-coupled magnetic beads

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S protein RBD MBS-K002 SARS-CoV-2 SARS-CoV-2 Spike protein RBD-coupled magnetic beads

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S1 protein SP-12 SARS-CoV-2 SARS-CoV-2 Spike S1 protein-Coated Plates, Clear, 96-Well

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S protein RBD RP-13 SARS-CoV-2 SARS-CoV-2 Spike protein RBD-Coated Plates, Clear, 96-Well

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Antibody
Molecule Cat. No. Species Product Description Protocol/Order
S protein RBD SAD-S35 Human Anti-SARS-CoV-2 neutralizing antibody (Human IgG1)

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Proteins
Molecule Cat. No. Species Host Product Description Protocol/Order
ACE2 AC2-H82E6 Human HEK293 Biotinylated Human ACE2 / ACEH Protein, His,Avitag™ (MALS verified)

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AC2-H82F9 Human HEK293 Biotinylated Human ACE2 / ACEH Protein, Fc,Avitag™

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AC2-H5257 Human HEK293 Human ACE2 / ACEH Protein, Fc Tag (MALS verified)

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AC2-C52H7 Cynomolgus HEK293 Cynomolgus ACE2 / ACEH Protein, His Tag

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AC2-H52H8 Human HEK293 Human ACE2 / ACEH Protein, His Tag (MALS verified)

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AC2-R5246 Rat HEK293 Rat ACE2 / ACEH Protein, His Tag (MALS verified)

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AC2-M5248 Mouse HEK293 Mouse ACE2 / ACEH Protein, His Tag (MALS verified)

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AC2-P5248 Paguma larvata HEK293 Paguma larvata ACE2 / ACEH Protein, His Tag

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S protein SPN-C52H4 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein (R683A, R685A), His Tag

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SPN-C52H8 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein (R683A, R685A) , His Tag, active trimer (MALS verified)

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SPN-S52H5 SARS HEK293 SARS S protein (R667A), His Tag

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S1 protein S1N-C52H3 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S1 protein, His Tag

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S1N-C82E8 SARS-CoV-2 HEK293 Biotinylated SARS-CoV-2 (COVID-19) S1 protein, His,Avitag™ (MALS verified)

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S1N-S52H5 SARS HEK293 SARS S1 protein, His Tag (MALS verified)

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S1N-C52H4 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S1 protein, His Tag (MALS verified)

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S1N-C5255 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S1 protein, Fc Tag

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S1N-C5257 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S1 protein, Mouse IgG2a Fc Tag

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S2 protein S2N-C5254 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S2 protein, Mouse IgG2a Fc Tag

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S protein RBD SPD-C82E9 SARS-CoV-2 HEK293 Biotinylated SARS-CoV-2 (COVID-19) S protein RBD, His,Avitag™ (MALS verified)

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SPD-C5255 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD, Fc Tag (MALS verified)

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SPD-S52H6 SARS HEK293 SARS S protein RBD, His Tag (MALS verified)

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SPD-C52H3 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD, His Tag (MALS verified)

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SPD-C5259 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD, Mouse IgG2a Fc Tag

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SPD-S52H4 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD (V367F), His Tag

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SPD-S52H5 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD (N354D), His Tag

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SPD-S52H7 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD (W436R), His Tag

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SPD-S52H8 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD (R408I), His Tag

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SPD-S52H3 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD (N354D, D364Y), His Tag

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Nucleocapsid protein NUN-C51H9 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) Nucleocapsid protein, His Tag

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NUN-C5227 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) Nucleocapsid protein, His Tag

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NUN-C81Q6 SARS-CoV-2 E.coli Biotinylated SARS-CoV-2 (COVID-19) Nucleocapsid protein, His,Avitag™

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Envelope protein ENN-C5128 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) Envelope protein, His Tag

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Papain-like Protease PAE-C5148 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) Papain-like Protease Protein, His Tag

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NSP1 NS1-C51H7 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) NSP1 Protein, His Tag

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NSP7 NS7-C51H6 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) NSP7 Protein, His Tag

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NSP8 NS8-C5149 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) NSP8 Protein, His Tag

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NSP7 & NSP8 NS8-C5125 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) NSP7&NSP8 Protein, His Tag

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NSP16 & NSP10 NS0-C51W3 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) NSP16&NSP10 Heterodimer Protein, His Tag&Twin Strep Tag

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S1 protein S1N-C5256 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S1 protein (D614G), His Tag

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预计上线时间:6月5日

SIN-V52H3 HCoV-NL63 HEK293 HCoV-NL63 S1 protein, His Tag

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SIN-V52H4 HCoV-229E HEK293 HCoV-229E S1 protein, His Tag

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SIN-V52H5 HCoV-OC43 HEK293 HCoV-OC43 S1 protein, His Tag

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SIN-V52H6 HCoV-HKU1(isolate N5) HEK293 HCoV-HKU1(isolate N5) S1 protein, His Tag

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S1 protein CTD S1D-C52H3 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S1 protein CTD, His Tag

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S1 protein NTD S1D-C52H6 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S1 protein NTD, His Tag

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S protein RBD SPD-C52H4 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD (G476S), His Tag

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SPD-C52H5 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S protein RBD (V483A), His Tag

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S2 protein S2N-C5253 SARS-CoV-2 HEK293 SARS-CoV-2 (COVID-19) S2 protein, Fc Tag

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预计上线时间:6月4日

Envelope protein ENN-C51P3 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) Envelope protein, His,BBP Tag

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预计上线时间:6月12日

NSP15 NS5-C5145 SARS-CoV-2 E.coli SARS-CoV-2 (COVID-19) NSP15 Protein, His Tag

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预计上线时间:6月15日


ACRO开发的高亲和力(经SPR/BLI/ELISA验证)的病毒相关蛋白希望能给抗病毒药物的开发提供有力的支持!

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