Abstract
BackgroundImmunoassays that employ multiplexed bead arrays produce high information content per sample. Such assays are now frequently used to evaluate humoral responses in clinical trials. Integrated software is needed for the analysis, quality control, and secure sharing of the high volume of data produced by such multiplexed assays. Software that facilitates data exchange and provides flexibility to perform customized analyses (including multiple curve fits and visualizations of assay performance over time) could increase scientists’ capacity to use these immunoassays to evaluate human clinical trials.ResultsThe HIV Vaccine Trials Network and the Statistical Center for HIV/AIDS Research and Prevention collaborated with LabKey Software to enhance the open source LabKey Server platform to facilitate workflows for multiplexed bead assays. This system now supports the management, analysis, quality control, and secure sharing of data from multiplexed immunoassays that leverage Luminex xMAP® technology. These assays may be custom or kit-based. Newly added features enable labs to: (i) import run data from spreadsheets output by Bio-Plex Manager™ software; (ii) customize data processing, curve fits, and algorithms through scripts written in common languages, such as R; (iii) select script-defined calculation options through a graphical user interface; (iv) collect custom metadata for each titration, analyte, run and batch of runs; (v) calculate dose–response curves for titrations; (vi) interpolate unknown concentrations from curves for titrated standards; (vii) flag run data for exclusion from analysis; (viii) track quality control metrics across runs using Levey-Jennings plots; and (ix) automatically flag outliers based on expected values. Existing system features allow researchers to analyze, integrate, visualize, export and securely share their data, as well as to construct custom user interfaces and workflows.ConclusionsUnlike other tools tailored for Luminex immunoassays, LabKey Server allows labs to customize their Luminex analyses using scripting while still presenting users with a single, graphical interface for processing and analyzing data. The LabKey Server system also stands out among Luminex tools for enabling smooth, secure transfer of data, quality control information, and analyses between collaborators. LabKey Server and its Luminex features are freely available as open source software at http://www.labkey.com under the Apache 2.0 license.
Highlights
Immunoassays that employ multiplexed bead arrays produce high information content per sample
Recent followup studies for the ALVAC-AIDSVAX trial, the first vaccine trial ever to demonstrate some degree of vaccine efficacy against human immunodeficiency virus 1 (HIV-1) [1,31], heavily used Luminex assays
Results include curve fit parameters, estimated concentrations interpolated from calculated curve fits for standard titrations, and metrics calculated from curve fits, including EC50, Area under the curve (AUC) and Highest mean fluorescence intensity (HighMFI)
Summary
Immunoassays that employ multiplexed bead arrays produce high information content per sample. Many [19,20,21,22,23] (but not all [24,25]) of these arrays leverage Luminex® xMAP® technology, so we refer to them here as “Luminex” assays These include both commercially available assay kits and custom assays, such as the binding complexity and dimensionality of such assays than ordinary ELISAs. The continued evolution of analysis and quality control techniques, the limits of current tools and the increasingly important role these assays play in certain fields of biomedical research all make enhanced software tools desirable for management and analysis of Luminex immunoassay data. Several of these studies used Luminex assays to examine the binding of plasma immunoglobulins to panels of HIV-1 envelope proteins to determine immune correlates of vaccine efficacy [1,5]
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