Highly efficient selection of phage antibodies mediated by display of antigen as Lpp-OmpA′ fusions on live bacteria 1 1Edited by J. Wells

Escherichia coli maltose-binding protein as a molecular chaperone for recombinant intracellular cytoplasmic single-chain antibodies 1 1Edited by R. Huber

“…We found that such crude extracts maintain most of the binding activity for several weeks of storage at 4 C with no appreciable degradation (not shown). A few examples include two anti ErbB2 scFvs that were isolated using DIP selection, 50 an additional anti b-galactosidase scFv, N9 (I.B., unpublished results) and the scFv of the anti myc 9E10 hybridoma. 51 A notable case is the anti¯uorescein Several MBP-scFvs were expressed in parallel in a trxB À isogenic strain of BL21(DE3) where cytoplasmic disul®de bond formation may occur to some extent.…”

“…E. coli b-galactosidase (Sigma, Israel) was coated at a concentration of 2 mg/ml. GST-ErbB2 fusionprotein 50 and FITC-albumin (Sigma, Israel) were coated at a concentration of 5 mg/ml. ScFvs or MBP-scFvs were applied onto the plates at various concentrations.…”

Peptide mimotopes recognized by antibodies cetuximab and matuzumab induce a functionally equivalent anti-EGFR immune response

“…Selection of peptide-displaying phage that interact with cetuximab and matuzumab Phage displaying peptides that specifically interact with cetuximab or matuzumab were selected by delayed infectivity panning (Benhar et al, 2000) from libraries of M13KE bacteriophage that display randomized linear 7mer, linear 12mer or cyclic 7mer peptides. For screening, we used noninfectable F À E. coli HB101 cells carrying single-chain antibody fragments (scFv) scFv(225) of cetuximab (Wels et al, 1995) or scFv(72000) of matuzumab as Lpp-OmpA' fusions (Georgiou et al, 1997) on the surface as schematically shown in Figures 1a and b.…”

“…For peptide selection, we used delayed infectivity panning (Benhar et al, 2000), which was based on the display of scFv antibody fragments of cetuximab and matuzumab on the surface of E. coli cells. This allowed rapid isolation of specific binders, and was expected to restrict selected peptides to those that either interact directly with the antigen binding sites of the antibodies encompassing the complementarity determining regions of heavy and light chain variable domains, or bind to adjacent framework regions that provide structural stability.…”

“…The peptides were selected from phage-displayed random peptide libraries by delayed infectivity panning (Benhar et al, 2000) using Escherichia coli cells that carry single-chain Fv (scFv) antibody fragments of cetuximab and matuzumab on their surface. Comparison of the selected peptides revealed several common sequence motifs not related to the primary amino-acid sequence of EGFR.…”