Figure 1.

Screening of VHH against NorC using yeast display. A, schematic of the yeast surface display strategy used for screening of VHH (ICab) binders against NorC (T4C) labeled with fluorescein (excitation, 494 nm; emission, 521 nm). A yeast display library built using vhh genes from PBMCs of an immunized camel was screened using anti-c-Myc antibody labeled with Alexa Fluor 647 (excitation, 650 nm; emission, 668 nm). B, FACS screening of uninduced yeast cells displays no shifts when incubated with fluorescein-labeled NorC and displays no nonspecific binding to anti-c-Myc antibody. Induced cells expressing ICab3 display diagonal shifts, signifying binding of NorC to surface-displayed ICab3.