Characterizing monoclonal antibody epitopes by filtered gene fragment phage display.
In the present paper, we describe a novel approach to map monoclonal antibody epitopes, using three new monoclonal antibodies that recognize h-TG2 (human transglutaminase 2) as an example. The target gene was fragmented and cloned upstream of an antibiotic-resistance gene, in the vector pPAO2, to select for in-frame polypeptides. After removal of the antibiotic-resistance gene by Cre/Lox recombination, an antigen fragment phage display library was created and selected against specific monoclonal antibodies. Using the h-TG2 fragment library, we were able to identify epitopes. This technique can also be broadly applied to the study of protein-protein interactions.
Di Niro
Roberto
R
Department of Biology, University of Trieste, Via Giorgieri 10, 34127 Trieste (TS), Italy.
Ferrara
Fortunato
F
Not
Tarcisio
T
Bradbury
Andrew R M
AR
Chirdo
Fernando
F
Marzari
Roberto
R
Sblattero
Daniele
D
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D004351:Drug Resistance / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology*; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000276:immunology; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D019151:Peptide Library; D011485:Protein Binding; D011487:Protein Conformation; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000502:physiology; D011503:Transglutaminases / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D019151:Peptide Library; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins
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