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Table 3 Phage panning on hFAP coated immunotubes and cells overexpressing mFAP

From: Species-Crossreactive scFv Against the Tumor Stroma Marker “Fibroblast Activation Protein” Selected by Phage Display From an Immunized FAP−/− Knock-Out Mouse

Panning Round

Enrichment Factor

Percent Binders

I. hFAP coated antigen

≥8#

25

II. mFAP+ cells

7

ND

III. mFAP+ cells

100

70

  1. Panning was carried out on recombinant human FAP coated on immunotubes (coated with 20 µg/tube) or on HEK 293 cells overexpressing mFAP (round II, 106 cells; round III, 105 cells in a total volume of 1 ml). In parallel, panning was done on noncoated tubes or control cells, respectively. To determine the enrichment factor, the phage output titer from panning on antigen was divided by the output titer of the control.
  2. Percentage of binders was determined after ELISA screening using CD8-hFAP fusion protein.
  3. #Plates of negative control showed no colonies
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Molecular Medicine

ISSN: 1528-3658