Expression and purification of a human pluripotent stem cell-specific lectin probe, rBC2LCN

Abstract

rBC2LCN is a recombinant N-terminal domain of BC2L-C lectin that is derived from the gram-negative bacteria Burkholderia cenocepacia and specifically binds to Fucα1-2Galβ1-3GlcNAc/GalNAc. Glycome analysis using a high-density lectin microarray revealed that rBC2LCN specifically binds to human pluripotent stem cells (hPSCs) but not to non-hPSCs. The lectin can be added to the cell culture medium for the live staining of hPSCs without causing visible cytotoxicity. Moreover, it can be used in flow cytometric analysis and for the staining of fixed hPSCs. rBC2LCN is a single-chain molecule with a low molecular weight (trimer of 16 kDa), which can be produced in large quantities in Escherichia coli (0.1 g/L). Therefore, rBC2LCN may be a cost-effective probe for use in hPSCs, unlike other hPSC surface marker antibodies that require a mammalian cell expression system for production. In this study, we describe the protocols for the expression and purification of rBC2LCN from E. coli and live staining of hPSCs using this probe.