Genetic and functional characterization of human pemphigus vulgaris monoclonal autoantibodies isolated by phage display
J. Clin. Invest. Aimee S. Payne, et al. 115:888 doi:10.1172/JCI24185 [Go to this article.]

Figure 1
Isolation of monovalent scFv mAbs. (A) ScFv nomenclature. (D3), (D1), or (D31) indicates the antigens used to select anti-Dsg mAbs from the scFv phage display library (Dsg3, Dsg1, or both, respectively). This is followed by a unique heavy chain and light chain nucleotide sequence designation. For mAbs that share the same clonal origin (same VDJ rearrangement) but differ in sequence because of somatic hypermutation, lower-case letters indicate unique members of a given clone (see Discussion and legend to Figure 9). (B) Soluble scFv mAbs were purified by nickel-chelation chromatography (see Methods). Two representative scFvs are shown by Coomassie blue staining after SDS-PAGE. (C) Gel-filtration HPLC demonstrates that scFvs are primarily monomeric in solution. Transferrin (Tf, 75 kDa) and carbonic anhydrase (CA, 30 kDa) served as protein standards, with 0.1 M Mops as a marker of the total volume of the column. The percentage of monomeric protein is shown, calculated from the area under the curve.