结构专家又添新作：经抗体验证的RSV Pre & Post F蛋白

转自：ACROBiosystems官方

不久前，Science公布了2022年度十大科学突破，RSV疫苗的突破进展成功入选。呼吸道合胞病毒（Respiratory Syncytial Virus, RSV）作为引起急性下呼吸道感染（ALRTIs）的主要原因之一，每年导致全球约3.31亿人发生呼吸道感染。然而，就是这样一个常见且早已被发现的呼吸道病毒，其疫苗的研发却十分坎坷，以至于至今仍无一款成熟的RSV疫苗问世。

此前的数十年间，科学家们不曾停止对于RSV疫苗的研究与开发，但均以失败告终。究其原因，可能是因为RSV病毒结构的特殊性。病毒表面的融合蛋白（fusion protein , F）和粘附蛋白（attachment protein, G）因在病毒入侵过程中的关键作用成为病毒研究的理想靶点，而其中G蛋白变异较多相对不稳定，F蛋白也存在Pre到Post构象转变影响其稳定性。

RSV F蛋白融合前及融合后构象

为了克服这一障碍，科学家们对RSV F蛋白的蛋白结构及其抗体进行了研究，发现该蛋白表面至少存在7个完全不同的抗原表位，并成功筛选到可以与不同表位结合的抗体，分别为φ site_D25/5C4/AM22，I site_131-2A/2F，II site_motavizumab/palivizumab，III site_MPE8，IV site_101F/mAb19，V site_hRSV90。其中识别IV表位的101F、识别II表位的palivizumab和识别I表位的131-2A抗体可以同时识别F蛋白的pre和post构象，识别φ表位的D25抗体只与Pre构象的F蛋白发生结合，识别III表位的MPE8抗体则偏好识别Pre构象，与Post构象具有较弱的结合活性[1]。2010年，Kwakkenbos研究团队筛选获得单克隆抗体AM14，其识别的抗原表位位于F-pre蛋白近赤道区域，横跨两个单体，可特异性识别F-pre三聚体结构，且被证明与此前中和效果最佳的D25抗体具有相似的中和活性和更快的动力学性质[2,3]。

RSV-F蛋白抗原表位及抗体结合情况[2,3]

ACROBiosystems，真正的蛋白结构专家👍

基于成熟的蛋白表达平台与结构验证方案，在对RSV病毒进行充分调研后，我们开发了RSV病毒研究关键的G蛋白与不同构象的F蛋白，并从上述抗体中选择了代表性抗体对所表达的Pre及Post构象F蛋白进行验证，力求为广大客户提供真正的Pre & Post RSV-F蛋白！

验证数据

★ 纯度经SDS-PAGE验证>95%，经SEC-MALS验证>90%。

The purity of HRSV (A) Fusion glycoprotein F0, His Tag is greater than 95% verified by SDS-PAGE and more than 90% verified by SEC-MALS.

★RSV Pre-F蛋白与Post-F蛋白和抗体101F、5C4、AM14、131-2A、D25结合活性经ELISA验证，与文献结果一致。

Immobilized HRSV (A) Post-fusion glycoprotein F0, His Tag and HRSV (A) Pre-fusion glycoprotein F0, His Tag at 1 μg/mL (100 μL/well) can bind Anti-Fusion glycoprotein F0 Antibody, Mouse IgG2a (101F) with a linear range of 0.1-4 ng/mL (QC tested).

Immobilized HRSV (A) Pre-fusion glycoprotein F0, His Tag at 1 μg/mL (100 μL/well) can bind Recombinant Mouse Anti-RSV F Antibody (5C4) with a linear range of 0.004-0.25 μg/mL. HRSV (A) Post-fusion glycoprotein F0, His Tag is verified not recoginized by Recombinant Mouse Anti-RSV F Antibody (5C4) (Routinely tested).

Immobilized HRSV (A) Pre-fusion glycoprotein F0, His Tag at 1 μg/mL (100 μL/well) can bind Recombinant Human Anti-RSV Antibody (AM14) with a linear range of 0.004-0.25 μg/mL. HRSV (A) Post-fusion glycoprotein F0, His Tag is verified not recognized by Recombinant Human Anti-RSV Antibody (AM14) (QC tested).

Immobilized HRSV (A) Post-fusion glycoprotein F0, His Tag and HRSV (A) Pre-fusion glycoprotein F0, His Tag at 1 μg/mL (100 μL/well) can bind Anti-RSV Fusion Protein Antibody (131-2A) with a linear range of 0.1-1 ng/mL . (Routinely tested).

Immobilized HRSV (A) Pre-fusion glycoprotein F0, His Tag at 1 μg/mL (100 μL/well) can bind Recombinant Human Anti-RSV Antibody (D25) with a linear range of 0.1-2 ng/mL. HRSV (A) Post-fusion glycoprotein F0, His Tag is verified not recoginized by Recombinant Human Anti-RSV Antibody (D25) in low concentration (Routinely tested).

参考文献：

1. McLellan JS, Chen M, Leung S, et al. Structure of RSV fusion glycoprotein trimer bound to a prefusion-specific neutralizing antibody. Science. 2013;340(6136):1113-1117. doi:10.1126/science.1234914

2. McLellan, J. S., Chen, M., Leung, S., Graepel, K. W., Du, X., Yang, Y., Zhou, T., Baxa, U., Yasuda, E., Beaumont, T., Kumar, A., Modjarrad, K., Zheng, Z., Zhao, M., Xia, N., Kwong, P. D., & Graham, B. S. (2013). Structure of RSV fusion glycoprotein trimer bound to a prefusion-specific neutralizing antibody. Science, 340(6136), 1113–1117. https://doi.org/10.1126/SCIENCE.1234914/SUPPL_FILE/MCLELLAN.SM.PDF

3. Gilman MSA, Moin SM, Mas V, Chen M, Patel NK, et al. (2015) Characterization of a Prefusion-Specific Antibody That Recognizes a Quaternary, Cleavage-Dependent Epitope on the RSV Fusion Glycoprotein. PLOS Pathogens 11(7): e1005035. https://doi.org/10.1371/journal.ppat.1005035

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