title: Monoclonal antibody proteomics: use of antibody mimotope displaying phages and the relevant synthetic peptides for mAb scouting.
abstract: Monoclonal antibody proteomics uses nascent libraries or cloned (Plasmascan™, QuantiPlasma™) libraries of mAbs that react with individual epitopes of proteins in the human plasma. At the initial phase of library creation, cognate protein antigen and the epitope interacting with the antibodies are not known. Scouting for monoclonal antibodies (mAbs) with the best binding characteristics is of high importance for mAb based biomarker assay development. However, in the absence of the identity of the cognate antigen the task represents a challenge. We combined phage display, and surface plasmon resonance (Biacore) experiments to test whether specific phages and the respective mimotope peptides obtained from large scale studies are applicable to determine key features of antibodies for scouting. We show here that mAb captured phage-mimotope heterogeneity that is the diversity of the selected peptide sequences, is inversely correlated with an important binding descriptor; the off-rate of the antibodies and that represents clues for driving the selection of useful mAbs for biomarker assay development. Carefully chosen synthetic mimotope peptides are suitable for specificity testing in competitive assays using the target proteome, in our case the human plasma.
authors: Targetex Biosciences, H-2120 Dunakeszi, Hungary. Electronic address: hajdu@target-ex.com.
authors: István
authors: I
authors: Hajdú
authors: Targetex Biosciences, H-2120 Dunakeszi, Hungary.
authors: Beáta
authors: B
authors: Flachner
authors: Targetex Biosciences, H-2120 Dunakeszi, Hungary.
authors: Melinda
authors: M
authors: Bognár
authors: Targetex Biosciences, H-2120 Dunakeszi, Hungary.
authors: Barbara M
authors: BM
authors: Végh
authors: Targetex Biosciences, H-2120 Dunakeszi, Hungary.
authors: Krisztina
authors: K
authors: Dobi
authors: Targetex Biosciences, H-2120 Dunakeszi, Hungary.
authors: Zsolt
authors: Z
authors: Lőrincz
authors: Biosystems International Kft., H-4032 Debrecen, Hungary.
authors: József
authors: J
authors: Lázár
authors: Targetex Biosciences, H-2120 Dunakeszi, Hungary.
authors: Sándor
authors: S
authors: Cseh
authors: Biosystems International Kft., H-4032 Debrecen, Hungary; Laboratory of Monoclonal Antibody Proteomics, Department of Human Genetics, University of Debrecen, H-4032, Hungary. Electronic address: laszlo.takacs@biosys-intl.com.
authors: László
authors: L
authors: Takács
authors: Biosystems International Kft., H-4032 Debrecen, Hungary.
authors: István
authors: I
authors: Kurucz
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D007118:Immunoassay; D007136:Immunoglobulins / Q000097:blood* / Q000235:genetics / Q000276:immunology; D007700:Kinetics; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D011485:Protein Binding; D040901:Proteomics / Q000379:methods*; D020349:Surface Plasmon Resonance
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D007136:Immunoglobulins; D019151:Peptide Library; D010455:Peptides
keywords: Affinity; Monoclonal antibody; Phage-display; Proteomics; Surface plasmon resonance