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

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Authors: Alexander E. Zambidis, Latha Kallur Siddaramaiah, A Konecny, Matthew D. Gray, Martin Prlic
Cited by: 0
References: 44

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.

Fields of Study & Subjects

Monoclonal and Polyclonal Antibodies Research Advanced Fluorescence Microscopy Techniques T-cell and B-cell Immunology

Publication details

Journal / source
bioRxiv (Cold Spring Harbor Laboratory)
Publication date
16 February 2026
Language
EN
Document type
Preprint