title: In vitro selection of other proteins than antibodies by means of ribosome display.
abstract: The in vitro selection and simultaneous evolution of proteins is feasible by means of ribosome display. Here, we describe the use of a protein bearing a binding property without being an antibody for affinity enrichment of the ternary complex, consisting of a protein, a ribosome and a encoding mRNA. The binding property was a simple affinity tag, namely Strep-tag II and His-tag. We could demonstrate that the selection of a specific mRNA encoding a shortened bovine heart fatty acid binding protein with a N-terminal His-tag was possible. After nine cycles of transcription, translation, affinity selection and reverse transcription PCR the protein with the His-tag could be enriched 10(8)-fold.
authors.affiliation: Institut für Biochemie, Freie Universität Berlin, Thielallee 63, D-14195 Berlin, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Lamla
authors.forename: V A
authors.initials: VA
authors.lastname: Erdmann
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics*; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; D016133:Polymerase Chain Reaction; D011691:Puromycin / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
chemical_list: D000906:Antibodies; D002352:Carrier Proteins; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D011691:Puromycin