title: Large scale production of phage antibody libraries using a bioreactor.
abstract: One of the limitations of the use of phage antibody libraries in high throughput selections is the production of sufficient phage antibody library at the appropriate quality. Here, we successfully adapt a bioreactor-based protocol for the production of phage peptide libraries to the production of phage antibody libraries. The titers obtained in the stirred-tank bioreactor are 4 to 5 times higher than in a standard shake flask procedure, and the quality of the phage antibody library produced is indistinguishable to that produced using standard procedures as assessed by Western blotting and functional selections. Availability of this protocol will facilitate the use of phage antibody libraries in high-throughput scale selections.
authors.affiliation: a New Mexico Consortium ; Los Alamos ; NM , USA.
authors.forename: Fortunato
authors.initials: F
authors.lastname: Ferrara
authors.forename: Chang-Yub
authors.initials: CY
authors.lastname: Kim
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Naranjo
authors.forename: Andrew R M
authors.initials: AR
authors.lastname: Bradbury
mesh_terms: D019149:Bioreactors; D019151:Peptide Library; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
chemical_list: D019151:Peptide Library; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies
keywords: antibody phage library production; bioreactor; phage display; scFv, single chain Fv; VH, Variable Heavy; VL, Variable Light; g3p, gene 3 protein; tTG2, tissue transglutaminase 2