Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: Plasmids used in this study.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Plasmid Preparation, Selection, Negative Control

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: Biophysical properties of the scFv purified in this study.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Purification

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: (A) western blot showing lane 1—binding of scFv with trastuzumab CDRs to the extracellular part of HER2 and lane 2—control sample showing binding of scFv with trastuzumab CDRs to the anti-His-tag secondary antibody; (a) trastuzumab + CyDisCo, (b) trastuzumab − CyDisCo, (c) trastuzumab CA − CyDisCo, (d) natalizumab framework-trastuzumab CDRs + CyDisCo, (e) Maa48 framework-trastuzumab CDRs + CyDisCo; (B) analysis of full length ITGA4 (purchased from 1. Abnova and 2. OriGene) on non-reducing SDS-PAGE; (C) dot blot of scFv containing natalizumab CDRs binding to full length ITGA4; (D) representative example of surface plasmon resonance (Biacore) analysis of binding of parental scFv IgA 1 to β-tubulin peptide epitope.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Western Blot, Binding Assay, Control, SDS Page, Dot Blot, SPR Assay

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: K D determination for scFv with trastuzumab CDRs binding to HER2 fragment V529-P625.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: K D determination for scFv with IgA 1 CDRs binding to β-tubulin peptide.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: (A) natalizumab framework—trastuzumab CDRs hybrid scFv. (B) natalizumab scFv. (C) trastuzumab scFv. For panels A-C all scFv were made in the presence of CyDisCo. The insert panel in each case shows the derivative of the change in fluorescence signal. (D) Correlation between melting temperature (Tm) and the yield of purified scFv expressed with (●) or without (○) disulfide bonds. Only those scFv showing a single thermal denaturation step are plotted. Coefficient of correlation between Tm and the yield of scFv with disulfide bonds R 2 = 0.12 and without disulfide bonds R 2 = 0.56.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Fluorescence, Purification

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: Thermal stability of the scFv.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Expressing