Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Endo-chip laser-induced thrombus formation: a vessel-on-chip model for in vitro testing of antithrombotic agents

doi: 10.1016/j.bvth.2025.100096

Figure Lengend Snippet: Inhibition of thrombus formation by inhibitory antiplatelet antibody 7E3 (abciximab) but not by low dose aspirin in the Endo-chip laser-injury model. (A) Blood was incubated with vehicle, aspirin (9 μg/mL), or abciximab (20 μg/mL) for 1 hour at RT. Blood was then labeled with anti-CD41 antibody and antifibrin antibody, recalcified with CaCl 2 (10 mM), and then immediately perfused through the Endo-chip at 100 s –1 , followed by laser injury to the endothelial layer. Representative images of platelet (CD41, green) and fibrin (magenta) accumulation at the injury site at 10 minutes. (B) Kinetics of platelet (upper panel) and fibrin (lower panel) surface area coverage over time. (C) Platelet (upper panel) and fibrin (lower panel) AUC after treatment of blood with vehicle, aspirin, or abciximab. Mean ± SD, n = 3 to 5, 1-way analysis of variance with Dunnett’s multiple comparisons test. AUC, area under the curve.

Article Snippet: Abciximab (anti-αIIbβ3 inhibitory antibody, clone C7E3) was purchased from MedChemExpress LLC (Monmouth Junction, NJ).

Techniques: Inhibition, Incubation, Labeling

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Cryo-Electron Microscopy Structure of the αIIbβ3–Abciximab Complex

doi: 10.1161/ATVBAHA.119.313671

Figure Lengend Snippet: Cryo-EM structure of the αIIbβ3–abciximab complex. (a) Density map of the αIIbβ3–abciximab complex at 2.8-Å resolution, segmented and colored according to the individual polypeptide chains. (b) Left panel: Atomic model of the αIIbβ3–abciximab complex colored as in panel (a). The green spheres represent the metal ions in the MIDAS, ADMIDAS and SyMBS. VH: variable domain of the heavy chain; CH1: first constant domain of the heavy chain; VL: variable domain of the light chain; CL: constant domain of the light chain. Hinge region denotes the flexible linkers that connect the variable and constant domains in abciximab. Right panel: Interaction surfaces of abciximab (top) and αIIbβ3 (bottom). Side chains of residues that are within 4.5 Å from a side chain of the interacting protein are shown in stick representation. (c) Fibrinogen/RGD-binding pocket in the cryo-EM structure of the αIIbβ3–abciximab complex (colored as in panel (a)) and in the X-ray crystal structure of the unliganded αIIbβ3 integrin (light grey with metal ions shown as black spheres). The overlay shows that abciximab binding does not induce meaningful differences in the binding pocket. The boxed area is shown in panel d. (d) Conformation of the β3 SDL loop in the cryo-EM structure of the αIIbβ3–abciximab complex (dark grey with SDL in blue) and in the X-ray crystal structure of the unliganded αIIbβ3 integrin (light grey with SDL in cyan). Abciximab is shown as an orange transparent surface to illustrate that the SDL in the conformation seen in the X-ray crystal structure of the unliganded αIIbβ3 integrin (cyan) would clash with abciximab.

Article Snippet: Protein sequencing The primary amino-acid sequence of abciximab was determined by de novo peptide sequencing 23 , 24 using tandem mass spectrometry (MS) by Rapid Novor Inc.

Techniques: Cryo-EM Sample Prep, Binding Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Cryo-Electron Microscopy Structure of the αIIbβ3–Abciximab Complex

doi: 10.1161/ATVBAHA.119.313671

Figure Lengend Snippet: Abciximab prevents ligands from access to the integrin ligand-binding pocket. (a) Shown are superimpositions of our structure of the αIIbβ3–abciximab complex (αIIbβ3 in grey ribbon representation and abciximab as transparent grey surface) with other ligand–integrin complexes, which are αVβ6 (salmon) in complex with pro-TGF-β (red) (PDB: 5FFO)38, αVβ3 (light blue) in complex with the tenth domain of fibronectin (dark green) (PDB: 4MMX)37, and αIIbβ3 (light green) in complex with a peptide of the fibrinogen γ-chain (blue) (PDB: 2VDO)35. The overlays show that ligand binding would cause sterical clashes with the bound abciximab. (b) Ligand binding has little effect on the abciximab-binding region of αIIbβ3. Left panel: Superimposition of our αIIbβ3–abciximab complex structure (αIIbβ3 shown in light grey and abciximab in yellow and orange) with the crystal structure of unbound αIIbβ3 (light green) (PDB: 3FCS)22 shows that abciximab does not affect the ligand-binding pocket. Right panel: Superimposition of our αIIbβ3–abciximab complex structure (αIIbβ3 shown in light grey and abciximab in yellow and orange) with the crystal structure of αIIbβ3 (dark green) in complex with a peptide of the fibrinogen γ-chain (red) (PDB: 2VDO)35 shows that peptide binding induces only subtle changes to the abciximab-interacting region of αIIbβ3.

Article Snippet: Protein sequencing The primary amino-acid sequence of abciximab was determined by de novo peptide sequencing 23 , 24 using tandem mass spectrometry (MS) by Rapid Novor Inc.

Techniques: Ligand Binding Assay, Binding Assay

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Cryo-Electron Microscopy Structure of the αIIbβ3–Abciximab Complex

doi: 10.1161/ATVBAHA.119.313671

Figure Lengend Snippet: Visualization of residues in (a) αIIbβ3 and (b) abciximab involved in inter-molecular contacts in at least 10% of the MD-simulation frames (see also Supplemental Figures VII and VIII). Insets in panel a refer to residues with calculated free-energy differences (ΔΔG(bind)) from wild type that are larger than 3.5 kcal/mol for top stabilizing (in green) and destabilizing (in purple) mutations of αIIbβ3 residues (see also Supplemental Figure IX).

Article Snippet: Protein sequencing The primary amino-acid sequence of abciximab was determined by de novo peptide sequencing 23 , 24 using tandem mass spectrometry (MS) by Rapid Novor Inc.

Techniques:

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Cryo-Electron Microscopy Structure of the αIIbβ3–Abciximab Complex

doi: 10.1161/ATVBAHA.119.313671

Figure Lengend Snippet: Residues of αIIbβ3 whose fluctuations are most affected by complex formation with abciximab are shown in stick representation and color-coded based on the change in RMSF. The RMSF changes are statistically significant in all of the residues depicted in stick representation (see also Supplemental Figure XI).

Article Snippet: Protein sequencing The primary amino-acid sequence of abciximab was determined by de novo peptide sequencing 23 , 24 using tandem mass spectrometry (MS) by Rapid Novor Inc.

Techniques: