Protocol to identify and monitor key mutations of broadly neutralizing antibody lineages following sequential immunization of Ig-humanized mice.
Protocol to identify and monitor key mutations of broadly neutralizing antibody lineages following sequential immunization of Ig-humanized mice.
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DOI：
10.1016/j.xpro.2022.101180
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发表时间：
2022-03-18
期刊：
影响因子：
--
通讯作者：
Mascola JR
中科院分区：
文献类型：
--
作者：
Chen X;Schmidt SD;Duan H;Doria-Rose NA;Mascola JR
以vrc01类抗hiv -1广泛中和抗体（bnAbs）为例，我们描述了一种通过点诱变和抗体结合和中和实验来鉴定bnAb功能关键突变的方案。我们还描述了通过纵向抗体突变谱监测特定免疫原反应中关键突变如何产生的步骤，这对疫苗评估和设计至关重要。该方案可定制用于其他v基因特异性bnab和动物模型。有关使用和执行此配置文件的完整详细信息，请参阅。通过结合和中和试验鉴定抗体功能的关键突变一种快速抗体关键突变分析和突变谱可视化程序纵向突变谱鉴定诱导关键抗体突变的免疫原以在顺序免疫的igg人源化小鼠中诱导的vrc01类抗hiv -1广泛中和抗体（bnAbs）为例。我们描述了一个方案，以确定关键突变的bnAb功能的点诱变和抗体结合和中和试验。我们还描述了通过纵向抗体突变谱监测特定免疫原反应中关键突变如何产生的步骤，这对疫苗评估和设计至关重要。该方案可定制用于其他v基因特异性bnab和动物模型。
Using the VRC01-class of anti-HIV-1 broadly neutralizing antibodies (bnAbs) elicited in sequentially immunized Ig-humanized mice as an example, we describe a protocol to identify key mutations for bnAb function by point mutagenesis and antibody binding and neutralization assays. We also describe steps to monitor how the key mutations arise in response to specific immunogens, which is critical for vaccine evaluation and design, via longitudinal antibody mutation profiling. This protocol can be customized for other V-gene-specific bnAbs and animal models. For complete details on the use and execution of this profile, please refer to. Create germline revertants for every amino acid mutation of a target bnAb Identify key mutations for antibody function by binding and neutralization assays A procedure for fast antibody key mutation analysis and mutation profile visualization Longitudinal mutation profiling identifies immunogens inducing key antibody mutations Using the VRC01-class of anti-HIV-1 broadly neutralizing antibodies (bnAbs) elicited in sequentially immunized Ig-humanized mice as an example, we describe a protocol to identify key mutations for bnAb function by point mutagenesis and antibody binding and neutralization assays. We also describe steps to monitor how the key mutations arise in response to specific immunogens, which is critical for vaccine evaluation and design, via longitudinal antibody mutation profiling. This protocol can be customized for other V-gene-specific bnAbs and animal models.
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影响因子：
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作者：
Giudicelli, Veronique;Brochet, Xavier;Lefranc, Marie-Paule
通讯作者：
Lefranc, Marie-Paule
DOI：
10.1126/science.1175868
发表时间：
2009-11-20
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chen L;Kwon YD;Zhou T;Wu X;O'Dell S;Cavacini L;Hessell AJ;Pancera M;Tang M;Xu L;Yang ZY;Zhang MY;Arthos J;Burton DR;Dimitrov DS;Nabel GJ;Posner MR;Sodroski J;Wyatt R;Mascola JR;Kwong PD
通讯作者：
Kwong PD
影响因子：
32.4
作者：
Chen X;Zhou T;Schmidt SD;Duan H;Cheng C;Chuang GY;Gu Y;Louder MK;Lin BC;Shen CH;Sheng Z;Zheng MX;Doria-Rose NA;Joyce MG;Shapiro L;Tian M;Alt FW;Kwong PD;Mascola JR
通讯作者：
Mascola JR
DOI：
10.1126/science.1234150
发表时间：
2013-05-10
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Jardine J;Julien JP;Menis S;Ota T;Kalyuzhniy O;McGuire A;Sok D;Huang PS;MacPherson S;Jones M;Nieusma T;Mathison J;Baker D;Ward AB;Burton DR;Stamatatos L;Nemazee D;Wilson IA;Schief WR
通讯作者：
Schief WR
影响因子：
14.9
作者：
Ye J;Ma N;Madden TL;Ostell JM
通讯作者：
Ostell JM