Details

Autor(en) / Beteiligte

• Xu, Lihui
• Aha, Patti
• Gu, Ke
• Kuimelis, Robert G.
• Kurz, Markus
• Lam, Terence
• Lim, Ai Ching
• Liu, Hongxiang
• Lohse, Peter A.
• Sun, Lin
• Weng, Shawn
• Wagner, Richard W.
• Lipovsek, Dasa

Titel

Directed Evolution of High-Affinity Antibody Mimics Using mRNA Display

Ist Teil von

• Chemistry & biology, 2002-08, Vol.9 (8), p.933-942

Ort / Verlag

United States: Elsevier Ltd

Erscheinungsjahr

2002

Link zum Volltext

Quelle

EZB Electronic Journals Library

Beschreibungen/Notizen

• We constructed a library of >10 12 unique, covalently coupled mRNA-protein molecules by randomizing three exposed loops of an immunoglobulin-like protein, the tenth fibronectin type III domain ( 10Fn3). The antibody mimics that bound TNF-α were isolated from the library using mRNA display. Ten rounds of selection produced 10Fn3 variants that bound TNF-α with dissociation constants (K d) between 1 and 24 nM. After affinity maturation, the lowest K d measured was 20 pM. Selected antibody mimics were shown to capture TNF-α when immobilized in a protein microarray. 10Fn3-based scaffold libraries and mRNA-display allow the isolation of high-affinity, specific antigen binding proteins; potential applications of such binding proteins include diagnostic protein microarrays and protein therapeutics.