Fig. 6: Both N and C-terminal sequences of the P17 tag contribute to enhanced scFv solubility in SHuffle T7 cells. | Nature Communications

Fig. 6: Both N and C-terminal sequences of the P17 tag contribute to enhanced scFv solubility in SHuffle T7 cells.

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

Fig. 6

a Amino acid sequences of wt P17 tag (P17-WT) and its truncated mutants with various deletions of C- or N-terminal residues. Basic, acidic, and polar residues are shown in blue, red, and green, respectively. The putative α-helix from P17 residues 22–29 is denoted with a cylinder. b Representative western blot to determine the solubility of MA18/7-scFv-HA protein and its derivatives fused to P17-WT or the C-terminally truncated mutants P17-C8del and P17-C15del. c Semiquantitative determination of the solubility of MA18/7-scFv-HA protein and its derivatives from b by three independent experiments. d Fold change of solubility of MA18/7-scFv derivatives from b compared to untagged MA18/7-scFv-HA. e Representative western blot to determine the solubility of MA18/7-scFv-HA protein and its derivatives fused to P17-WT or the N-terminally truncated mutants P17-N8del, P17-N14del, and P17-N18del. f Semiquantitative determination of the solubility of MA18/7-scFv-HA protein and its derivatives from e by three independent experiments. g Fold change of solubility of MA18/7-scFv derivatives from e compared to untagged MA18/7-scFv-HA. T total protein, S soluble protein. Data are presented as mean values ± SD. The statistical significance of differences between the two experimental groups was assessed by one-way ANOVA.

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