SARS-CoV-2 Spike RBD Recombinant Nanobody

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說(shuō)明書(shū)

中文名稱：

SARS-CoV-2 Spike RBD重組納米抗體
貨號(hào)：

CSB-RA33245A2GMY
規(guī)格：

￥3080
圖片：
The Binding Activity of SARS-CoV-2 Spike RBD Nanobody with SARS-CoV-2-S1-RBD
Activity: Measured by its binding ability in a functional ELISA. Immobilized SARS-CoV-2-S1-RBD (CSB-YP3324GMY1) at 2 μg/ml can bind SARS-CoV-2 Spike RBD Nanobody, the EC50 is 0.8674 ng/ml.

In the Colloidal Gold Immunochromatography Assay detection system, the background of antibody (CSB-RA33245A2GMY) is clean, the detection limit can be as low as 25ng/ml (1.75ng/0.07ml), and the sensitivity is very good.

SARS-CoV-2 Spike RBD Nanobody (CSB-RA33245A2GMY) competed with ACE2-HRP conjugate (CSB-MP866317HU) for binding to SARS-CoV-2-S1-RBD (CSB-YP3324GMY1). The binding signal of SARS-CoV-2-S1-RBD and ACE2-HRP conjugate was gradually reduced as the SARS-CoV-2 Spike RBD Nanobody concentrations increased. It indicated that this SARS-CoV-2 Spike RBD Nanobody effectively inhibited the SARS-CoV-2-S1-RBD/ACE2 binding. And the IC₅₀ of this SARS-CoV-2 Spike RBD Nanobody is 1.296 nM.

SARS-CoV-2 Spike RBD Nanobody (CSB-RA33245A2GMY) competitively prevented SARS-CoV-2-S1-RBD (CSB-YP3324GMY1) from binding to ACE2-HRP conjugate (CSB-MP866317HU). The inhibition efficacy of the SARS-CoV-2-S1-RBD/ACE2 binding was positively proportionally to the SARS-CoV-2 Spike RBD Nanobody concentrations. It showed that this SARS-CoV-2 Spike RBD Nanobody effectively inhibited the SARS-CoV-2-S1-RBD/ACE2 binding. And the IC₅₀ of this SARS-CoV-2 Spike RBD Nanobody is 0.1074 μg/ml.

SARS-CoV-2 Spike protein RBD His/Sumostar Tag (CSB-YP3324GMY1) captured on COOH chip binding to the SARS-CoV-2 Spike RBD Nanobody (CSB-RA33245A2GMY), increases the local refractive index (RI), leading to a red shift of the LSPR peak position. The higher concentrations of SARS-CoV-2 Spike RBD Nanobody, the larger the wavelength shift. The detected affinity constant of SARS-CoV-2 Spike protein RBD/SARS-CoV-2 Spike RBD Nanobody binding is 28.2nM.

ELISA: Immobilize various types of SARS proteins at concentration of 2μg/ml on solid substrate, then react with SARS-CoV-2 Spike RBD Nanobody at concentration of 100μg/ml, 10μg/ml and 1μg/ml. It shows the SARS-CoV-2 Spike RBD Nanobody (CSB-RA33245A2GMY) is specific for SARS-CoV-2-S1-RBD protein, without any cross-reactivity with MERS-CoV, SARS-CoV, HCoV-OC43 or HCoV-229E.
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靶點(diǎn)詳情

產(chǎn)品詳情

產(chǎn)品描述：

本產(chǎn)品為針對(duì)SARS-CoV-2刺突蛋白受體結(jié)合域（Spike RBD）的重組納米抗體（CSB-RA33245A2GMY），專為新冠病毒相關(guān)研究設(shè)計(jì)。靶點(diǎn)Spike RBD是介導(dǎo)病毒入侵宿主細(xì)胞的核心功能域，其與ACE2受體的結(jié)合機(jī)制研究對(duì)理解病毒感染及開(kāi)發(fā)干預(yù)策略具有重要價(jià)值。經(jīng)多平臺(tái)驗(yàn)證，該抗體在ELISA檢測(cè)中靈敏度達(dá)1:100000稀釋度，膠體金免疫層析（GICA）應(yīng)用中可穩(wěn)定工作于1:40000稀釋條件，中和實(shí)驗(yàn)顯示在1:100至1:10000濃度梯度內(nèi)均能有效阻斷病毒侵染。適用于體外病毒中和效價(jià)評(píng)估、免疫檢測(cè)試劑的開(kāi)發(fā)優(yōu)化及抗病毒藥物篩選等科研場(chǎng)景。產(chǎn)品采用哺乳動(dòng)物表達(dá)系統(tǒng)制備，具有高親和力與批次穩(wěn)定性，其單域抗體結(jié)構(gòu)賦予其優(yōu)異的組織穿透性和耐高溫特性，特別適用于快速檢測(cè)試劑的凍干工藝開(kāi)發(fā)。本制劑經(jīng)內(nèi)毒素去除工藝處理，可直接用于細(xì)胞水平實(shí)驗(yàn)，為科研工作者研究新冠病毒侵染機(jī)制及開(kāi)發(fā)新型防治方案提供可靠工具。
Uniprot No.：

P0DTC2
別名：

S; 2; Spike glycoprotein; S glycoprotein; E2; Peplomer protein)
反應(yīng)種屬：

Human Novel Coronavirus (SARS-CoV-2/ 2019-nCoV)
免疫原：

Recombinant Human Novel Coronavirus Spike glycoprotein(S) (319-541aa) (CSB-YP3324GMY1 and CSB-MP3324GMY1b1)
免疫原種屬：

Human Novel Coronavirus (SARS-CoV-2/ 2019-nCoV)
標(biāo)記方式：

Non-conjugated
克隆類型：

Monoclonal
抗體亞型：

VHH fusion with human IgG1 Fc
純化方式：

Affinity-chromatography
克隆號(hào)：

A1
濃度：

It differs from different batches. Please contact us to confirm it.
保存緩沖液：

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
產(chǎn)品提供形式：

Liquid
應(yīng)用范圍：

ELISA, GICA, Neutralising

推薦稀釋比：

Application	Recommended Dilution
ELISA	1:10000-1:100000
GICA	1:10000-1:40000
Neutralising	1:100-1:10000

Protocols：

ELISA Protocol
儲(chǔ)存條件：

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
貨期：

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
用途：

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

產(chǎn)品評(píng)價(jià)

靶點(diǎn)詳情

功能：

attaches the virion to the cell membrane by interacting with host receptor, initiating the infection. Binding to human ACE2 receptor and internalization of the virus into the endosomes of the host cell induces conformational changes in the Spike glycoprotein. Binding to host NRP1 and NRP2 via C-terminal polybasic sequence enhances virion entry into host cell. This interaction may explain virus tropism of human olfactory epithelium cells, which express high level of NRP1 and NRP2 but low level of ACE2. The stalk domain of S contains three hinges, giving the head unexpected orientational freedom. Uses human TMPRSS2 for priming in human lung cells which is an essential step for viral entry. Can be alternatively processed by host furin. Proteolysis by cathepsin CTSL may unmask the fusion peptide of S2 and activate membranes fusion within endosomes.; mediates fusion of the virion and cellular membranes by acting as a class I viral fusion protein. Under the current model, the protein has at least three conformational states: pre-fusion native state, pre-hairpin intermediate state, and post-fusion hairpin state. During viral and target cell membrane fusion, the coiled coil regions (heptad repeats) assume a trimer-of-hairpins structure, positioning the fusion peptide in close proximity to the C-terminal region of the ectodomain. The formation of this structure appears to drive apposition and subsequent fusion of viral and target cell membranes.; Acts as a viral fusion peptide which is unmasked following S2 cleavage occurring upon virus endocytosis.; May down-regulate host tetherin (BST2) by lysosomal degradation, thereby counteracting its antiviral activity.
基因功能參考文獻(xiàn)：
1. Study presents crystal structure of C-terminal domain of SARS-CoV-2 (SARS-CoV-2-CTD) spike S protein in complex with human ACE2 (hACE2); hACE2-binding mode similar overall to that observed for SARS-CoV. However, details at the binding interface show that key residue substitutions in SARS-CoV-2-CTD slightly strengthen the interaction and lead to higher affinity for receptor binding than SARS-CoV receptor-binding domain. PMID: 32378705
2. crystal structure of the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 bound to the cell receptor ACE2 PMID: 32365751
3. crystal structure of the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 (engineered to facilitate crystallization) in complex with ACE2 PMID: 32320687
4. Out of the two isolates from India compared to the isolates from Wuhan, China, one was found to harbor a mutation in its receptor-binding domain (RBD) at position 407 where, arginine was replaced by isoleucine. This mutation has been seen to change the secondary structure of the protein at that region and this can potentially alter receptor binding of the virus. PMID: 32275855
5. Structural modeling of the SARS-CoV-2 spike glycoprotein show similar receptor utilization between SARS-CoV-2 and SARS-CoV, despite a relatively low amino acid similarity in the receptor binding module. Compared to SARS-CoV and all other coronaviruses in Betacoronavirus lineage B, an extended structural loop containing basic amino acids were identified at the interface of the receptor binding (S1) and fusion (S2) domains. PMID: 32245784
6. crystal structure of CR3022, a neutralizing antibody from a SARS patient, in complex with the receptor-binding domain of the SARS-CoV-2 spike (S) protein to 3.1 A; study provides insight into how SARS-CoV-2 can be targeted by the humoral immune response and revealed a conserved, but cryptic epitope shared between SARS-CoV-2 and SARS-CoV PMID: 32225176
7. SARS-CoV and SARS-CoV-2 spike proteins have comparable binding affinities achieved by balancing energetics and dynamics. The SARS-CoV-2-ACE2 complex contains a higher number of contacts, a larger interface area, and decreased interface residue fluctuations relative to the SARS-CoV-ACE2 complex. PMID: 32225175
8. Interaction interface between cat/dog/pangolin/Chinese hamster ACE2 and SARS-CoV/SARS-CoV-2 S protein was simulated through homology modeling. Authors identified that N82 of ACE2 showed closer contact with receptor-binding domain of S protein than human ACE2. PMID: 32221306
9. SARS-CoV-2 S glycoprotein harbors a furin cleavage site at the boundary between the S1/S2 subunits, which is processed during biogenesis and sets this virus apart from SARS-CoV and SARS-related CoVs; determined cryo-EM structures of the SARS-CoV-2 S ectodomain trimer. PMID: 32201080
10. Study demonstrates that SARS-CoV-2 uses the SARS-CoV receptor ACE2 for entry and the serine protease TMPRSS2 for S protein priming. PMID: 32155444
11. The ACE2-B0AT1 complex exists as a dimer of heterodimers. Structural alignment of the RBD-ACE2-B0AT1 ternary complex with the S protein of SARS-CoV-2 suggests that two S protein trimers can simultaneously bind to an ACE2 homodimer. PMID: 32142651
12. study demonstrated SARS-CoV-2 S protein entry on 293/hACE2 cells is mainly mediated through endocytosis, and PIKfyve, TPC2 and cathepsin L are critical for virus entry; found that SARS-CoV-2 S protein could trigger syncytia in 293/hACE2 cells independent of exogenous protease; there was limited cross-neutralization activity between convalescent sera from SARS and COVID-19 patients PMID: 32132184
13. study determined a 3.5-angstrom-resolution cryo-electron microscopy structure of the 2019-nCoV S trimer in the prefusion conformation; provided biophysical and structural evidence that the 2019-nCoV S protein binds angiotensin-converting enzyme 2 (ACE2) with higher affinity than does severe acute respiratory syndrome (SARS)-CoV S PMID: 32075877
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亞細(xì)胞定位：

Virion membrane; Single-pass type I membrane protein. Host endoplasmic reticulum-Golgi intermediate compartment membrane; Single-pass type I membrane protein. Host cell membrane; Single-pass type I membrane protein.
蛋白家族：

Betacoronaviruses spike protein family

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