title: Bacterial secretion of soluble and functional trivalent scFv-based N-terminal trimerbodies.
abstract: Recombinant antibodies are used with great success in many different diagnostic and therapeutic applications. A variety of protein expression systems are available, but nowadays almost all therapeutic antibodies are produced in mammalian cell lines due to their complex structure and glycosylation requirements. However, production of clinical-grade antibodies in mammalian cells is very expensive and time-consuming. On the other hand, Escherichia coli (E. coli) is known to be the simplest, fastest and most cost-effective recombinant expression system, which usually achieves higher protein yields than mammalian cells. Indeed, it is one of the most popular host in the industry for the expression of recombinant proteins. In this work, a trivalent single-chain fragment variable (scFv)-based N-terminal trimerbody, specific for native laminin-111, was expressed in human embryonic kidney 293 cells and in E. coli. Mammalian and bacterially produced anti-laminin trimerbody molecules display comparable functional and structural properties, although importantly the yield of trimerbody expressed in E. coli was considerably higher than in human cells. These results demonstrated that E. coli is a versatile and efficient expression system for multivalent trimerbody-based molecules that is suitable for their industrial production.
authors.affiliation: Department of Antibody Engineering, Leadartis S.L., Madrid, Spain, ana.blanco@leadartis.es.
authors.forename: Ana
authors.initials: A
authors.lastname: Blanco-Toribio
authors.forename: Ana
authors.initials: A
authors.lastname: Álvarez-Cienfuegos
authors.forename: Noelia
authors.initials: N
authors.lastname: Sainz-Pastor
authors.forename: Nekane
authors.initials: N
authors.lastname: Merino
authors.forename: Marta
authors.initials: M
authors.lastname: Compte
authors.forename: Laura
authors.initials: L
authors.lastname: Sanz
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Blanco
authors.forename: Luis
authors.initials: L
authors.lastname: Álvarez-Vallina