【热门产品推介】InVivoMAb Anti-HRSV F/F protein Antibody (3M3)

货号：VVV02804

产品链接：http://www.antibodysystem.com/product/13701.html

寄主物种：Human

物种反应性：Human respiratory syncytial virus (HRSV)

形式：Liquid

存储缓冲区：0.01M PBS, pH 7.4.

浓度：1 mg/ml

纯度：>95% by SDS-PAGE.

克隆性：Monoclonal

应用：ELISA, In vivo Neutralization

储存：Use a manual defrost freezer and avoid repeated freeze-thaw cycles.Store at +4°C short term (1-2 weeks).Store at -20 °C 12 months. Store at -80°C long term.

参考文献：

Hematopoietic PBX-interacting protein mediates cartilage degeneration during the pathogenesis of osteoarthritis. PMID: 30659184
Autophagy-based unconventional secretion of HMGB1 by keratinocytes plays a pivotal role in psoriatic skin inflammation. PMID: 32019420
Major vault protein (MVP) negatively regulates osteoclastogenesis via calcineurin-NFATc1 pathway inhibition. PMID: 34158848
Head-to-Head Comparison of Fibroblast Activation Protein Inhibitors (FAPI) Radiotracers versus [18F]F-FDG in Oncology: A Systematic Review. PMID: 34681850
"In-loop" 18 F-fluorination: A proof-of-concept study. PMID: 31083778
Estrogen Signals Through Peroxisome Proliferator-Activated Receptor-γ Coactivator 1α to Reduce Oxidative Damage Associated With Diet-Induced Fatty Liver Disease. PMID: 27658772
筛选并构建成功表达A亚型人呼吸道合胞病毒(Human Respiratory Syncytial Virus, HRSV)融合前构象F蛋白的重组人5型腺病毒(Recombinant Human type5 adenovirus, rHADV-5),为制备和研发HRSV重组腺病毒载体疫苗奠定基础。

方法 

在真核表达载体pcDNA3.1(+)上插入具有不同突变位点的HRSV F蛋白编码序列，利用融合前F蛋白单克隆抗体AM14和D25筛选HRSV F蛋白突变体；通过同源重组将筛选出的前构象F蛋白突变体编码序列插入非复制型人5型腺病毒载体，在HEK293细胞中进行重组病毒的包装和扩增，经氯化铯密度梯度法获得纯化的重组病毒；应用夹心ELISA法和Western blot鉴定F蛋白的表达及构象。

结果 

通过AM14和D25单克隆抗体筛选出了2个稳定维持在融和前构象的HRSV F蛋白序列(SC-3M和SC-5M)。将SC-3M和SC-5M编码序列插入非复制型人5型腺病毒载体，对两种重组人5型腺病毒(pAd5-SC-3M和pAd5-SC-5M)和空载体(pAd5-vector)纯化，病毒滴度分别达到2.016×10~(10),2.52×10~(10)和2.948×10~(11) IU/mL。pAd5-SC-3M和pAd5-SC-5M感染HEK293后均能高效表达HRSV融合前构象的F蛋白并维持三聚体结构，且pAd5-SC-5M感染细胞中F蛋白表达量高于pAd5-SC-3M。

结论 

成功在体外传代细胞中获得可高效表达A型HRSV融合前F蛋白的重组人5型腺病毒，为进一步通过动物试验评价该HRSV重组腺病毒载体疫苗奠定了基础。