CaptureBody enables accurate unmixing for spectral flow cytometry.

Accurate spectral unmixing is a critical step for flow cytometry data analysis and requires a single stain control for every fluorescent parameter used in an experiment. Currently, compensation/unmixing particles are often used for making single stain controls when a target protein is of low abundance or a cell type is of low frequency. However, compensation/unmixing particles introduce incongruencies in emission spectra, compared to cells, resulting in spectral unmixing or compensation errors. To enable the use of cells regardless of the abundance of target proteins or immune cell type, we generated a bispecific antibody that links a human anti-CD45 and mouse anti-immunoglobulin G (IgG) variable region. We refer to this bispecific tool as CaptureBody (CB) and highlight the benefits of its final nanobody-based design. We provide all sequences and methods necessary for the in-house expression of a CaptureBody in order to disseminate their use for spectral flow cytometry experiments.