Single-chain antibodies for the conformation-specific blockade of activated platelet integrin  IIb  3 designed by subtractive selection from naïve human.

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Single-chain antibodies for the conformation-specific blockade of activated platelet integrin  IIb  3 designed by subtractive selection from naïve human phage libraries Supplemental files Supplemental Fig. I: Binding of scFv-Clones to recombinant CHO cells. Flow cytometric histograms of scFvs (2  g/ml) binding on CHO cells expressing wild type  IIb  3 (gray histograms) and “activated” (VGFFK-deleted)  IIb  3 (white histograms) CHO cells for three representative clones (MA2, SCE5 and MA4). Supplemental Fig. II: Expression level of CHO cells transfected with wild type  IIb  3 (“wild type” CHO cells) and VGFFK-deleted and thus activated  IIb  3 (“VGGFK-deleted CHO cells). Flow cytometric histograms as obtained with specific antibodies against the two subunits of  IIb  3, CD 41 (second row) and CD61 (third row) and an isotypic control (first row). Supplemental Fig III: Exclusion of crossreactivity with other integrins. Depicted are the histograms obtained in flow cytometry with scFv clones MA4, MA2 and SCE5 (2  g/ml) on recombinant CHO cells expressing “activated” (VGFFK-deleted) Mac-1 and “activated” (VGFFK-deleted) vitronectin receptor  V  3.

Supplemental Fig I: Binding of scFv clones to recombinant CHO cells

Non-expressing CHO cells „Wild type“ CHO cells „VGFFK-deleted“ CHO cells Supplemental Fig. II: Expression level of recombinant CHO cells

CHO Mac-1 Del aVb3 Del Supplemental Fig III: Exclusion of crossreactivity with other integrins control scFv MA4 scFv MA2 scFv SCE5