LabKey Software, Seattle, Washington 98102, USA.
BMC Immunol. 2011 May 27;12:33. doi: 10.1186/1471-2172-12-33.
Multiple types of assays allow sensitive detection of virus-specific neutralizing antibodies. For example, the extent of antibody neutralization of HIV-1, SIV and SHIV can be measured in the TZM-bl cell line through the degree of luciferase reporter gene expression after infection. In the past, neutralization curves and titers for this standard assay have been calculated using an Excel macro. Updating all instances of such a macro with new techniques can be unwieldy and introduce non-uniformity across multi-lab teams. Using Excel also poses challenges in centrally storing, sharing and associating raw data files and results.
We present LabKey Server's NAb tool for organizing, analyzing and securely sharing data, files and results for neutralizing antibody (NAb) assays, including the luciferase-based TZM-bl NAb assay. The customizable tool supports high-throughput experiments and includes a graphical plate template designer, allowing researchers to quickly adapt calculations to new plate layouts. The tool calculates the percent neutralization for each serum dilution based on luminescence measurements, fits a range of neutralization curves to titration results and uses these curves to estimate the neutralizing antibody titers for benchmark dilutions. Results, curve visualizations and raw data files are stored in a database and shared through a secure, web-based interface. NAb results can be integrated with other data sources based on sample identifiers. It is simple to make results public after publication by updating folder security settings.
Standardized tools for analyzing, archiving and sharing assay results can improve the reproducibility, comparability and reliability of results obtained across many labs. LabKey Server and its NAb tool are freely available as open source software at http://www.labkey.com under the Apache 2.0 license. Many members of the HIV research community can also access the LabKey Server NAb tool without installing the software by using the Atlas Science Portal (https://atlas.scharp.org). Atlas is an installation of LabKey Server.
多种类型的检测方法可用于敏感地检测病毒特异性中和抗体。例如，可以通过感染后荧光素酶报告基因表达的程度，在 TZM-bl 细胞系中测量 HIV-1、SIV 和 SHIV 的抗体中和程度。过去，该标准检测的中和曲线和滴度是使用 Excel 宏计算的。使用新技术更新此类宏的所有实例可能很麻烦，并且会导致多实验室团队之间的不一致性。使用 Excel 还会在集中存储、共享和关联原始数据文件和结果方面带来挑战。
我们介绍了 LabKey Server 的 NAb 工具，用于组织、分析和安全共享中和抗体（NAb）检测数据、文件和结果，包括基于荧光素酶的 TZM-bl NAb 检测。该可定制工具支持高通量实验，包括图形板模板设计器，允许研究人员快速适应新板布局的计算。该工具根据发光测量值计算每个血清稀释度的中和百分比，拟合一系列中和曲线以得出滴定结果，并使用这些曲线估算基准稀释度的中和抗体滴度。结果、曲线可视化和原始数据文件存储在数据库中，并通过安全的基于 Web 的界面共享。可以根据样品标识符将 NAb 结果与其他数据源集成。发布后，通过更新文件夹安全设置，可以轻松公开结果。
用于分析、存档和共享检测结果的标准化工具可以提高许多实验室获得的结果的可重复性、可比性和可靠性。LabKey Server 及其 NAb 工具可在 http://www.labkey.com 上免费获得，可根据 Apache 2.0 许可证获得开源软件。许多 HIV 研究界成员也可以通过使用 Atlas Science Portal（https://atlas.scharp.org）访问 LabKey Server NAb 工具，而无需安装软件。Atlas 是 LabKey Server 的一个安装。
