title: A highly functional synthetic phage display library containing over 40 billion human antibody clones.
abstract: Several synthetic antibody phage display libraries have been created and used for the isolation of human monoclonal antibodies. The performance of antibody libraries, which is usually measured in terms of their ability to yield high-affinity binding specificities against target proteins of interest, depends both on technical aspects (such as library size and quality of cloning) and on design features (which influence the percentage of functional clones in the library and their ability to be used for practical applications). Here, we describe the design, construction and characterization of a combinatorial phage display library, comprising over 40 billion human antibody clones in single-chain fragment variable (scFv) format. The library was designed with the aim to obtain highly stable antibody clones, which can be affinity-purified on protein A supports, even when used in scFv format. The library was found to be highly functional, as >90% of randomly selected clones expressed the corresponding antibody. When selected against more than 15 antigens from various sources, the library always yielded specific and potent binders, at a higher frequency compared to previous antibody libraries. To demonstrate library performance in practical biomedical research projects, we isolated the human antibody G5, which reacts both against human and murine forms of the alternatively spliced BCD segment of tenascin-C, an extracellular matrix component frequently over-expressed in cancer and in chronic inflammation. The new library represents a useful source of binding specificities, both for academic research and for the development of antibody-based therapeutics.
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zürich, Zürich, Switzerland; Philochem AG, Otelfingen, Switzerland.
authors.forename: Marcel
authors.initials: M
authors.lastname: Weber
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zürich, Zürich, Switzerland; Philochem AG, Otelfingen, Switzerland.
authors.forename: Emil
authors.initials: E
authors.lastname: Bujak
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zürich, Zürich, Switzerland; Philochem AG, Otelfingen, Switzerland.
authors.forename: Alessia
authors.initials: A
authors.lastname: Putelli
authors.affiliation: Philochem AG, Otelfingen, Switzerland.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Villa
authors.affiliation: Philochem AG, Otelfingen, Switzerland.
authors.forename: Mattia
authors.initials: M
authors.lastname: Matasci
authors.affiliation: Philochem AG, Otelfingen, Switzerland.
authors.forename: Laura
authors.initials: L
authors.lastname: Gualandi
authors.affiliation: Philochem AG, Otelfingen, Switzerland.
authors.forename: Teresa
authors.initials: T
authors.lastname: Hemmerle
authors.affiliation: Philochem AG, Otelfingen, Switzerland.
authors.forename: Sarah
authors.initials: S
authors.lastname: Wulhfard
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zürich, Zürich, Switzerland.
authors.forename: Dario
authors.initials: D
authors.lastname: Neri
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000918:Antibody Specificity; D003001:Cloning, Molecular; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D019063:Tenascin / Q000737:chemistry / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D019151:Peptide Library; D019063:Tenascin