INVESTIGATING NEUTRALIZING ANTIBODY INDUCED CONFORMATIONAL CHANGES AND MATURATIO

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 先前的SAXS对已知结构的控制蛋白和几种gp140三聚体形式的结果表明，SAXS是一种易于掌握的分析方法。所有样品都可以在短时间内准备好，用于SAXS数据收集，目前正在进行常规生产，以支持其他研究。蛋白质的单分散性由SEC通过同时进行静态/动态光散射以及各种形式的定量/定性抗体结合分析来监测。我们希望获得不同gp140-NAB复合体的整体形状和尺寸信息，如果可能的话，为包膜尖峰的功能形式建立模型，并通过环境尖峰的冷冻-EM结构以及gp120和gp41的晶体结构进行验证。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Prior SAXS results with control proteins of known structure and several gp140 trimer forms show that SAXS is a tractable method of analysis in our hands. All samples can be prepared on short notice for SAXS data collection and are currently in routine production to support other studies. Protein monodispersity is monitored by SEC with simultaneous static/dynamic light scattering as well as quantitative/qualitative Ab binding assays in a variety of formats. We expect to obtain overall shape and size information of different gp140-NAb complexes, generate models for the functional form of envelope spike if possible, and verify by cryo-EM structures of Env spikes and crystal structures of gp120 and gp41.