title: A fully synthetic human Fab antibody library based on fixed VH/VL framework pairings with favorable biophysical properties.
abstract: This report describes the design, generation and testing of Ylanthia, a fully synthetic human Fab antibody library with 1.3E+11 clones. Ylanthia comprises 36 fixed immunoglobulin (Ig) variable heavy (VH)/variable light (VL) chain pairs, which cover a broad range of canonical complementarity-determining region (CDR) structures. The variable Ig heavy and Ig light (VH/VL) chain pairs were selected for biophysical characteristics favorable to manufacturing and development. The selection process included multiple parameters, e.g., assessment of protein expression yield, thermal stability and aggregation propensity in fragment antigen binding (Fab) and IgG1 formats, and relative Fab display rate on phage. The framework regions are fixed and the diversified CDRs were designed based on a systematic analysis of a large set of rearranged human antibody sequences. Care was taken to minimize the occurrence of potential posttranslational modification sites within the CDRs. Phage selection was performed against various antigens and unique antibodies with excellent biophysical properties were isolated. Our results confirm that quality can be built into an antibody library by prudent selection of unmodified, fully human VH/VL pairs as scaffolds.
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Tiller
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Schuster
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Dorothée
authors.initials: D
authors.lastname: Deppe
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Siegers
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Strohner
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Herrmann
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Marion
authors.initials: M
authors.lastname: Berenguer
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Dominique
authors.initials: D
authors.lastname: Poujol
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Stehle
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Stark
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Heßling
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Daubert
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Felderer
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Kaden
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Johanna
authors.initials: J
authors.lastname: Kölln
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Enzelberger
authors.affiliation: MorphoSys AG; Martinsried/Planegg, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Urlinger
mesh_terms: D000911:Antibodies, Monoclonal / Q000235:genetics / Q000627:therapeutic use*; D000915:Antibody Affinity; D002478:Cells, Cultured; D022801:Complementarity Determining Regions / Q000235:genetics; D019281:Dimerization; D015195:Drug Design; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007167:Immunotherapy; D015202:Protein Engineering; D055550:Protein Stability
chemical_list: D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains
keywords: CDR-H3 design; Slonomics; VH/VL pairing; Ylanthia; antibody engineering; human antibody library; phage display