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Figure 2 | Microbial Cell Factories

Figure 2

From: Bacterial cytoplasm as an effective cell compartment for producing functional VHH-based affinity reagents and Camelidae IgG-like recombinant antibodies

Figure 2

Comparison between antibody constructs expressed in the bacterial cytoplasm and periplasm. a) The affinity of the VHH A10 for its antigen HER2 (purified ectodomain) was measured by Surface Plasmon Resonance. Three constructs were compared: A10 tagged myc + His and expressed in the bacterial periplasm, A10 tagged SNAP + His and A10 tagged SORT + His, both expressed in the bacterial cytoplasm. The graphs correspond to a single experiment representative of at least three repeats. The analyte concentrations were in the range 300-3.5 nM (myc-tag), 100-3.5 nM (SNAP-tag), and 30-0.35 nM (SORT-tag). b) The anti-HER2 antibodies A10 fused to GFP was expressed in the cytoplasm and directly assessed by FACS (filter band-pass: 515-545 nm) using HER2 negative (MCF10A) and positive (SKBR3) cells. c) The anti-HER2 antibodies A10 and C8 were expressed fused to the LPTEG peptide (SORT tag) in both the periplasm and the cytoplasm and the resulting samples were assessed by FACS (filter band-pass: 564-606 nm) using HER2 negative (MCF10A) and positive (SKBR3) cells, and the 6xHis tag for visualization.

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Microbial Cell Factories

ISSN: 1475-2859

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