CaptureBody – an anti-CD45 x anti-IgG bispecific antibody 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 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 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-IgG variable region. We refer to this new 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 to disseminate their use for spectral flow cytometry experiments.

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Publication Details

Journal
bioRxiv (Cold Spring Harbor Laboratory)
Published
2026-02-16
DOI
https://doi.org/10.64898/2026.02.13.704926
Primary Topic
Single-cell and spatial transcriptomics
Type
article
Field-Weighted Citation Impact
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article

CaptureBody – an anti-CD45 x anti-IgG bispecific antibody enables accurate unmixing for spectral flow cytometry

Latha Kallur Siddaramaiah, Martin Prlic, A Konecny, Alexander E. Zambidis
bioRxiv (Cold Spring Harbor Laboratory)
Single-cell and spatial transcriptomics
article

CaptureBody – an anti-CD45 x anti-IgG bispecific antibody enables accurate unmixing for spectral flow cytometry

Latha Kallur Siddaramaiah, Martin Prlic, A Konecny, Alexander E. Zambidis
article en

Abstract

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 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 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-IgG variable region. We refer to this new 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 to disseminate their use for spectral flow cytometry experiments.

bioRxiv (Cold Spring Harbor Laboratory)
University of Washington (US), Fred Hutch Cancer Center (US)
Openalex Percentile: Top 52%
Single-cell and spatial transcriptomics
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