Figure 1
From: IgG-single-chain TRAIL fusion proteins for tumour therapy
Structure and basic biochemical properties of IgG-scTRAIL fusion proteins. (a) Scheme of single-chain TRAIL fusion to the C-terminus of the anti-EGFR IgG1 heavy and light chain. Underlined aa residues at both termini of the TRAIL subunits indicate substitutions compared to wild-type TRAIL. CH2 and CH3 harbour mutations which abrogate ADCC/CDC functionality. A 15 aa residues peptide linker comprising a sequence motif with two N-glycosylation sites (AAGNGTSNGTSEFGG) was used for genetic fusion of CH3 or CL domains with the first subunit of single-chain TRAIL. L, leader peptide; F, FLAG tag. (b) Drawings of full IgG-scTRAIL fusion proteins depicting control anti-EGFR huC225 IgG1 (hu225 IgG), fusion of scTRAIL to hu225 IgG light chains (LC-scTRAIL), fusion of scTRAIL to hu225 IgG heavy chains (HC-scTRAIL) and fusion of scTRAIL to both hu225 IgG chains (LC/HC-scTRAIL). (c) Hu225 IgG and scTRAIL fusion variants (5 µg each) were analysed by non-reducing (left) and reducing (right) SDS-PAGE and subsequent Coomassie staining. scT, single-chain TRAIL. (d) Hu225 IgG and scTRAIL fusion variants and were analysed by size exclusion chromatography under native buffer conditions. Elution times and molecular masses of globular reference proteins are indicated by dotted lines.
