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: Yields in mg/L/OD 600 of scFv expressed with CyDisCo (black bars) or without (white bars). Average OD 600 of the cultures was 22 ± 3 for those containing CyDisCo and 27 ± 3 for cultures without CyDisCo. Error bars show standard deviation, n = 3–4. scFv frameworks which have all four cysteines mutated to alanine are indicated as CA.

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: Standard Deviation

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: Relative yield of scFv expressed without CyDisCo compared to the yield from expression with CyDisCo, n = 3–4.

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

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: CD spectra of scFv that were found to be soluble in the absence of disulfide bonds and comparisons with the disulfide bonded counterpart. (A) scFv trastuzumab; (B) scFv natalizumab; (C) scFv trastuzumab and natalizumab and the trastuzumab framework—natalizumab CDR hybrid scFv all produced with CyDisCo; (D) trastuzumab framework—natalizumab CDR hybrid scFv. scFv frameworks which have all four cysteines mutated to alanine are indicated as CA.

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: Circular Dichroism, Produced

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

Journal: Cureus

Article Title: Exacerbation of Paraneoplastic Neurologic Syndrome After Immune Checkpoint Inhibitor Treatment in Advanced Small Cell Lung Cancer: A Case Report

doi: 10.7759/cureus.79272

Figure Lengend Snippet: Clinical characteristics, authors, and published literature of 27 cases of paraneoplastic neurological syndrome associated with small-cell lung cancer exacerbated by immune checkpoint inhibitors combined with previously reported cases and the present case. CY: Cyclophosphamide; GC: Glucocorticoids; IVIg: Intravenous immunoglobulin therapy; PE: Plasma exchange; SSN: Subacute sensory neuropathy *For those who had symptoms before ICI treatment, the period was summarized from the introduction of ICIs until the worsening of symptoms, and for those who had no symptoms before ICI treatment, the period was summarized from the introduction of ICIs until the appearance of symptoms.

Article Snippet: 19 , 71 , F , Nivolumab + ipilimumab , Short-term memory impairment , Hu , NO , After 4 days , GC, natalizumab , Partly improved , Hottinger et al. 2018 , [ ] .

Techniques: Clinical Proteomics