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. 1992 Mar 15;89(6):2429–2433. doi: 10.1073/pnas.89.6.2429

Directed evolution of a protein: selection of potent neutrophil elastase inhibitors displayed on M13 fusion phage.

B L Roberts 1, W Markland 1, A C Ley 1, R B Kent 1, D W White 1, S K Guterman 1, R C Ladner 1
PMCID: PMC48671  PMID: 1549606

Abstract

Inhibitors of human neutrophil elastase were engineered by designing and producing a library of phage-displayed protease inhibitory domains derived from wild-type bovine pancreatic trypsin inhibitor and fractionating the library for binding to the target protease. The affinity of one of the engineered variants for human neutrophil elastase (Kd = 1.0 pM) is 3.6 x 10(6)-fold higher than that of the parental protein and exceeds the highest affinity reported for any reversible human neutrophil elastase inhibitor by 50-fold. Thus the display phage method has allowed us to obtain protein derivatives that exhibit greatly increased affinity for a predetermined target. The technology can be applied to design high-affinity proteins for a wide variety of target molecules.

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Selected References

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