貨號：VVV02804

產(chǎn)品鏈接：http://www.antibodysystem.com/product/13701.html

寄主物種：Human

物種反應(yīng)性：Human respiratory syncytial virus (HRSV)

形式：Liquid

存儲緩沖區(qū)：0.01M PBS, pH 7.4.

濃度：1 mg/ml

純度：>95% by SDS-PAGE.

克隆性：Monoclonal

應(yīng)用：ELISA, In vivo Neutralization

儲存：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.

參考文獻(xiàn)：

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 in?ammation. 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
篩選并構(gòu)建成功表達(dá)A亞型人呼吸道合胞病毒(Human Respiratory Syncytial Virus, HRSV)融合前構(gòu)象F蛋白的重組人5型腺病毒(Recombinant Human type5 adenovirus, rHADV-5),為制備和研發(fā)HRSV重組腺病毒載體疫苗奠定基礎(chǔ)。

方法?

在真核表達(dá)載體pcDNA3.1(+)上插入具有不同突變位點(diǎn)的HRSV F蛋白編碼序列，利用融合前F蛋白單克隆抗體AM14和D25篩選HRSV F蛋白突變體；通過同源重組將篩選出的前構(gòu)象F蛋白突變體編碼序列插入非復(fù)制型人5型腺病毒載體，在HEK293細(xì)胞中進(jìn)行重組病毒的包裝和擴(kuò)增，經(jīng)氯化銫密度梯度法獲得純化的重組病毒；應(yīng)用夾心ELISA法和Western blot鑒定F蛋白的表達(dá)及構(gòu)象。

結(jié)果?

通過AM14和D25單克隆抗體篩選出了2個(gè)穩(wěn)定維持在融和前構(gòu)象的HRSV F蛋白序列(SC-3M和SC-5M)。將SC-3M和SC-5M編碼序列插入非復(fù)制型人5型腺病毒載體，對兩種重組人5型腺病毒(pAd5-SC-3M和pAd5-SC-5M)和空載體(pAd5-vector)純化，病毒滴度分別達(dá)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后均能高效表達(dá)HRSV融合前構(gòu)象的F蛋白并維持三聚體結(jié)構(gòu)，且pAd5-SC-5M感染細(xì)胞中F蛋白表達(dá)量高于pAd5-SC-3M。

結(jié)論?

成功在體外傳代細(xì)胞中獲得可高效表達(dá)A型HRSV融合前F蛋白的重組人5型腺病毒，為進(jìn)一步通過動物試驗(yàn)評價(jià)該HRSV重組腺病毒載體疫苗奠定了基礎(chǔ)。