title: Improved stability of multivalent antibodies containing the human collagen XV trimerization domain.
abstract: We recently described the in vitro and in vivo properties of an engineered homotrimeric antibody made by fusing the N-terminal trimerization region of collagen XVIII NC1 domain to the C-terminus of a scFv fragment [trimerbody (scFv-NC1) 3; 110 kDa]. Here, we demonstrated the utility of the N-terminal trimerization region of collagen XV NC1 domain in the engineering of trivalent antibodies. We constructed several scFv-based trimerbodies containing the human type XV trimerization domain and demonstrated that all the purified trimerbodies were trimeric in solution and exhibited excellent antigen binding capacity. Importantly, type XV trimerbodies demonstrated substantially greater thermal and serum stability and resistance to protease digestion than type XVIII trimerbodies. In summary, the small size, high expression level, solubility and stability of the trimerization domain of type XV collagen make it the ideal choice for engineering homotrimeric antibodies for cancer detection and therapy.
authors.affiliation: Madrid, Spain; Molecular Immunology Unit; Hospital Universitario Puerta de Hierro.
authors.forename: Angel M
authors.initials: AM
authors.lastname: Cuesta
authors.affiliation: Madrid, Spain; Molecular Immunology Unit; Hospital Universitario Puerta de Hierro.
authors.forename: David
authors.initials: D
authors.lastname: Sánchez-Martín
authors.affiliation: Madrid, Spain; Leadartis S.L., Ferraz 3.
authors.forename: Ana
authors.initials: A
authors.lastname: Blanco-Toribio
authors.affiliation: Structural Biology Unit; CIC bioGUNE, Parque Tecnológico de Bizkaia; Derio, Spain.
authors.forename: Maider
authors.initials: M
authors.lastname: Villate
authors.affiliation: Madrid, Spain; Molecular Immunology Unit; Hospital Universitario Puerta de Hierro.
authors.forename: Kelly
authors.initials: K
authors.lastname: Enciso-Álvarez
authors.affiliation: Madrid, Spain; Leadartis S.L., Ferraz 3.
authors.forename: Ana
authors.initials: A
authors.lastname: Alvarez-Cienfuegos
authors.affiliation: Madrid, Spain; Molecular Immunology Unit; Hospital Universitario Puerta de Hierro.
authors.forename: Noelia
authors.initials: N
authors.lastname: Sainz-Pastor
authors.affiliation: Madrid, Spain; Molecular Immunology Unit; Hospital Universitario Puerta de Hierro.
authors.forename: Laura
authors.initials: L
authors.lastname: Sanz
authors.affiliation: Structural Biology Unit; CIC bioGUNE, Parque Tecnológico de Bizkaia; Derio, Spain; IKERBASQUE; Basque Foundation for Science; Bilbao, Spain.
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Blanco
authors.affiliation: Madrid, Spain; Molecular Immunology Unit; Hospital Universitario Puerta de Hierro.
authors.forename: Luis
authors.initials: L
authors.lastname: Alvarez-Vallina
mesh_terms: D000818:Animals; D000912:Antibodies, Neoplasm / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D003094:Collagen / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000276:immunology / Q000473:pathology; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D000912:Antibodies, Neoplasm; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D003094:Collagen