Author/Authors :
Lihui Xu، نويسنده , , Patti Aha، نويسنده , , Ke Gu، نويسنده , , Robert G. Kuimelis، نويسنده , , Markus Kurz، نويسنده , , Terence Lam، نويسنده , , Elison Ai Ching Lim، نويسنده , , Hongxiang Liu، نويسنده , , Peter A. Lohse، نويسنده , , Lin Sun، نويسنده , , Shawn Weng، نويسنده , , Richard W. Wagner، نويسنده , , Dasa Lipovsek، نويسنده ,
Abstract :
We constructed a library of >1012 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 (Kd) between 1 and 24 nM. After affinity maturation, the lowest Kd 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.
