mab1864  (R&D Systems)

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet: Production time course of lassotide 35 in four fed-batch fermenters (5 L working volume). Reactors were fed based on control of dissolved oxygen (DO) which was maintained at 20%. Production was shown to increase at 32 hours, when cell growth decreased. Average titers over 4 tanks are plotted ± SEM. See Experimental Methods and for further details.

Article Snippet: For integrin α5β1, binding was visualized using a mouse anti-human integrin a5 (CD49e) monoclonal antibody that targets the α5 subunit (cat. no. MAB1864-SP was purchased from R&D Systems).

Techniques: Control

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet: Production time course of lassotide 35 over 72 h in 4 x 5 L fermentations. B . Associated cell growth over time, as measured by OD600 through 72 h in 4 fermenters. Titers for tanks 1-4 (U5-U8) were 5.39, 5.64, 5.70, and 5.96 g/L, respectively. SEM was calculated as 0.10 g/L from these values.

Article Snippet: For integrin α5β1, binding was visualized using a mouse anti-human integrin a5 (CD49e) monoclonal antibody that targets the α5 subunit (cat. no. MAB1864-SP was purchased from R&D Systems).

Techniques:

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet: In vitro metabolic stability and plasma protein binding for lassotide 36 and conjugated derivative 47 . A . Plasma stability data, as measured by incubating 36 and 47 in human and mouse plasma. B. Hepatocyte metabolism stability data, as measured by incubating 36 and 47 with cryopreserved human and mouse hepatocytes. C. Plasma protein binding (PPB) data as measured by rapid equilibrium dialysis. Experiments were performed in duplicate, and data are presented as mean values. Lassotide concentrations = 2 μM in all studies. See Experimental Methods section for details.

Article Snippet: For integrin α5β1, binding was visualized using a mouse anti-human integrin a5 (CD49e) monoclonal antibody that targets the α5 subunit (cat. no. MAB1864-SP was purchased from R&D Systems).

Techniques: In Vitro, Clinical Proteomics, Protein Binding

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet:

Article Snippet: For integrin α5β1, binding was visualized using a mouse anti-human integrin a5 (CD49e) monoclonal antibody that targets the α5 subunit (cat. no. MAB1864-SP was purchased from R&D Systems).

Techniques: Software

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet: Analytical QC check for lassotides purified by preparative HPLC. Purified lassotides were analyzed using a Shimadzu LC-10ADvp HPLC equipped with a Vydac C18 25 cm x 4.6 mm, 5-micron column. Mobile phase was 0.1% TFA in 15-20% CH3CN increased to 35-50% CH3CN over 20 min. A . Lassotide 35 , B . Lassotide 36 , C . Lassotide 38 , D . Lassotide 47 . In all cases, lassotide purity was ≥ 98% in all cases.

Article Snippet: For integrin α5β1, binding was visualized using a mouse anti-human integrin a5 (CD49e) monoclonal antibody that targets the α5 subunit (cat. no. MAB1864-SP was purchased from R&D Systems).

Techniques: Purification

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet: Production time course of lassotide 35 in four fed-batch fermenters (5 L working volume). Reactors were fed based on control of dissolved oxygen (DO) which was maintained at 20%. Production was shown to increase at 32 hours, when cell growth decreased. Average titers over 4 tanks are plotted ± SEM. See Experimental Methods and for further details.

Article Snippet: To each well, 50 mL of 0.5 μg/mL anti-α5 antibody (R&D Systems, cat. no. MAB1864-SP) was added and incubated for 1 hour at room temperature.

Techniques: Control

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet: Production time course of lassotide 35 over 72 h in 4 x 5 L fermentations. B . Associated cell growth over time, as measured by OD600 through 72 h in 4 fermenters. Titers for tanks 1-4 (U5-U8) were 5.39, 5.64, 5.70, and 5.96 g/L, respectively. SEM was calculated as 0.10 g/L from these values.

Article Snippet: To each well, 50 mL of 0.5 μg/mL anti-α5 antibody (R&D Systems, cat. no. MAB1864-SP) was added and incubated for 1 hour at room temperature.

Techniques:

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet: In vitro metabolic stability and plasma protein binding for lassotide 36 and conjugated derivative 47 . A . Plasma stability data, as measured by incubating 36 and 47 in human and mouse plasma. B. Hepatocyte metabolism stability data, as measured by incubating 36 and 47 with cryopreserved human and mouse hepatocytes. C. Plasma protein binding (PPB) data as measured by rapid equilibrium dialysis. Experiments were performed in duplicate, and data are presented as mean values. Lassotide concentrations = 2 μM in all studies. See Experimental Methods section for details.

Article Snippet: To each well, 50 mL of 0.5 μg/mL anti-α5 antibody (R&D Systems, cat. no. MAB1864-SP) was added and incubated for 1 hour at room temperature.

Techniques: In Vitro, Clinical Proteomics, Protein Binding

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet:

Article Snippet: To each well, 50 mL of 0.5 μg/mL anti-α5 antibody (R&D Systems, cat. no. MAB1864-SP) was added and incubated for 1 hour at room temperature.

Techniques: Software

Journal: bioRxiv

Article Title: Overcoming Immune Checkpoint Inhibitor Resistance via Potent and Selective Dual αvβ6/8 Inhibitors Based on Engineered Lasso Peptides

doi: 10.1101/2025.01.28.635346

Figure Lengend Snippet: Analytical QC check for lassotides purified by preparative HPLC. Purified lassotides were analyzed using a Shimadzu LC-10ADvp HPLC equipped with a Vydac C18 25 cm x 4.6 mm, 5-micron column. Mobile phase was 0.1% TFA in 15-20% CH3CN increased to 35-50% CH3CN over 20 min. A . Lassotide 35 , B . Lassotide 36 , C . Lassotide 38 , D . Lassotide 47 . In all cases, lassotide purity was ≥ 98% in all cases.

Article Snippet: To each well, 50 mL of 0.5 μg/mL anti-α5 antibody (R&D Systems, cat. no. MAB1864-SP) was added and incubated for 1 hour at room temperature.

Techniques: Purification