Fig. 1: P17 tag enhanced solubility of four scFvs in E. coli SHuffle T7 cells. | Nature Communications

Fig. 1: P17 tag enhanced solubility of four scFvs in E. coli SHuffle T7 cells.

From: A 33-residue peptide tag increases solubility and stability of Escherichia coli produced single-chain antibody fragments

Fig. 1

a Determination of dilution ranges of total and soluble fractions of G12-scFv-HA protein for linear correlation with western blotting signals. Upper panel: ScFvs from 5 μl total and soluble bacterial protein samples serially diluted with 2× SDS-PAGE loading buffer were detected by western blotting using an anti-His-tag. The relative density value (rel. DV) of each full-length (FL) scFv was obtained by setting the density of total scFv diluted by 1/5 at 100%. Middle panel: Over the entire dilution range rel. DV of both total and soluble scFvs correlated with dilutions logarithmically as calculated by the non-linear fitting program of Origin 8.0 software. Bottom panel: In the dilution range from 1/20 to 1/160, rel. DV of both total and soluble scFvs correlated linearly with dilutions as fulfilled by a linear fitting program of Origin 8.0. Linear equations of rel. DV and dilution (D) are also presented. b Determination of solubility of G12-scFv-HA and G12-scFv-HA-P17 proteins expressed in E. coli SHuffle T7 and BL21(DE3) Star strains. ScFv’s solubility was determined as the percentage ratio of soluble versus total scFvs after their dilutions were normalized at 1/160, and the result of one representative experiment is shown. − below the detection limit, T total protein, S soluble protein. c Determination of solubility of VRC01-scFv-HA, ADRI-scFv-HA, MA18/7-scFv-HA, and their P17-tagged derivatives expressed in SHuffle T7 strain. The result of one representative experiment was shown. d Fold change of solubility of P17-tagged scFvs to corresponding untagged scFvs and the isoelectric point (pI) of each untagged scFv.

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