Skip to main content
. 2018 May 29;13:3189–3201. doi: 10.2147/IJN.S164542

Figure 1.

Figure 1

Expression and purification of S-Fab from E. coli.

Notes: (A) The bacterial S-Fab expression constructs contain a pelB signal sequence, anti-CD3 (human UCTH1 clone) VH (or VL) and CH1 (CL), and anti-CEA-VHH. To facilitate antibody detection and purification, a flag-tag and his6-tag were added to the C-terminal end of the heavy and light chains, respectively. (B) Schematic representation of S-Fab after co-expression. (C) Coomassie blue-stained SDS-PAGE chromatogram of purified S-Fab after the two-step purification. + indicates reducing condition (2 μM 2-mecaptoethanol); - indicates nonreducing condition (no 2-mecaptoethanol). (D) Gel filtration analysis showing that the molecular weight of S-Fab was ~130 kDa. M (kDa), molecular weight markers (kilodalton).

Abbreviations: CEA, carcinoembryonic antigen; E. coli, Escherichia coli; S-Fab, single-domain antibody-linked Fab; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; UV, ultraviolet.