Label-Free Primary Screening And Affinity Ranking Of Fragment Libraries Using Parallel Analysis Of Protein Panels  

Oleh:

Hämäläinen, Markku D. ; Zhukov, Andrei ; Ivarsson, Maria ; Fex, Tomas ; Gottfries, Johan ; Karlsson, Robert

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Concurrent Engineering vol. 13 no. 3 (Sep. 2005), page 202-209.
Topik: label-free screening; fragment library; protein; high-throughput screening
Fulltext: 202.pdf (891.23KB)

Isi artikel

The authors present fragment screening data obtained using a label-free parallel analysis approach where the binding of fragment library compounds to 4 different target proteins can be screened simultaneously using surface plasmon resonance detection. They suggest this method as a first step in fragment screening to identify and select binders, reducing the demanding requirements on subsequent X-ray or nuclear magnetic resonance studies, and as a valuable “clean-up” tool to eliminate unwanted promiscuous binders from libraries. A small directed fragment library of known thrombin binders and a general 500-compound fragment library were used in this study. Thrombin, blocked thrombin, carbonic anhydrase, and glutathione-Stransferase were immobilized on the sensor chip surface, and the direct binding of the fragments was studied in real time. Only 12 µg of each protein is needed for screening of a 3000-compound fragment library. For screening, a binding site-blocked target as reference facilitates the identification of binding site-selective hits and the signals from other reference proteins for the elimination of false positives. The scope and limitations of this screening approach are discussed for both target-directed and general fragment libraries. (Journal of Biomolecular Screening 2008:202-209)

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