Sensitive Protein Detection Using Site-Specifically Oligonucleotide-Conjugated Nanobodies.

ABSTRACT: High-quality affinity probes are critical for sensitive and specific protein detection, in particular for detection of protein biomarkers in the early phases of disease development. Proximity extension assays (PEAs) have been used for high-throughput multiplexed protein detection of up to a few thousand different proteins in one or a few microliters of plasma. Clonal affinity reagents can offer advantages over the commonly used polyclonal antibodies (pAbs) in terms of reproducibility and standardization of such assays. Here, we explore nanobodies (Nbs) as an alternative to pAbs as affinity reagents for PEA. We describe an efficient site-specific approach for preparing high-quality oligo-conjugated Nb probes via enzyme coupling using Sortase A (SrtA). The procedure allows convenient removal

SUBMITTER: Al-Amin RA 

PROVIDER: S-EPMC9310004 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature