Modular DNA barcoding of nanobodies enables multiplexed in situ protein imaging and high-throughput biomolecule detection.

ABSTRACT: Current immunodetection methods using antibody-DNA conjugates enable multiplexed target detection through orthogonal DNA barcodes, but existing conjugation approaches are labor-intensive and often compromise antibody function. Here, we present a modular, site-specific, and cost-efficient DNA tagging strategy - multiplexed and modular barcoding of antibodies (MaMBA). Utilizing nanobodies as modular adaptors, MaMBA enables direct site-specific labeling of off-the-shelf IgG antibodies with a one-component design. We first applied MaMBA to develop the MaMBA-assisted immunosignal hybridization chain reaction (misHCR) method for highly multiplexed in situ protein imaging via orthogonal HCR. Its cleavable variant, misHCRⁿ, achieves simultaneous visualization of 12 differe

SUBMITTER: Zhong S 

PROVIDER: S-EPMC12283080 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature