Single-chain antibodies for the conformation-specific blockade of

activated platelet integrin IIb3 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 IIb3 (gray

histograms) and “activated” (VGFFK-deleted) IIb3 (white histograms) CHO

cells for three representative clones (MA2, SCE5 and MA4).

Supplemental Fig. II:

Expression level of CHO cells transfected with wild type IIb3 (“wild type” CHO

cells) and VGFFK-deleted and thus activated IIb3 (“VGGFK-deleted CHO

cells). Flow cytometric histograms as obtained with specific antibodies against

the two subunits of IIb3, 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 V3.

 

His Tag MA4MA2 SCE5

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