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Multivalent avimer proteins evolved by exon shuffling of a family of human receptor domains

A Corrigendum to this article was published on 01 February 2006

Abstract

We have developed a class of binding proteins, called avimers, to overcome the limitations of antibodies and other immunoglobulin-based therapeutic proteins. Avimers are evolved from a large family of human extracellular receptor domains by in vitro exon shuffling and phage display, generating multidomain proteins with binding and inhibitory properties. Linking multiple independent binding domains creates avidity and results in improved affinity and specificity compared with conventional single-epitope binding proteins. Other potential advantages over immunoglobulin domains include simple and efficient production of multitarget-specific molecules in Escherichia coli, improved thermostability and resistance to proteases. Avimers with sub-nM affinities were obtained aganist five targets. An avimer that inhibits interleukin 6 with 0.8 pM IC50 in cell-based assays is biologically active in two animal models.

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Figure 1: Overview of avimer technology.
Figure 2: Avimer properties.
Figure 3: Anti-IL-6 Avimer characterization.

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Acknowledgements

The authors would like to thank Evan Green, Cynthia Bailey, Kevin Allen, Anielka Montalvan and Khue Dang for their expert laboratory assistance, Jeff Smith and Peter van Vlasselaer for their assistance in overall project management, all the contributors at Boehringer-Ingleheim Austria for their work in fermentation and production scale-up, and Harry Ruan of Comparative Biosciences for performing the in vivo portion of the experiment shown in Figure 3.

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Correspondence to Joshua Silverman or Willem Pim C Stemmer.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Alignment of human A-domain sequences. (PDF 130 kb)

Supplementary Fig. 2

Pharmacokinetics of an avimer in cynomolgus. (PDF 55 kb)

Supplementary Fig. 3

Surface plasmon resonance analysis of C326 binding to IL-6 immobilized to a CM-5 chip. (PDF 118 kb)

Supplementary Fig. 4

Avimers are not immunogenic in mice. (PDF 81 kb)

Supplementary Data (PDF 26 kb)

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Silverman, J., Lu, Q., Bakker, A. et al. Multivalent avimer proteins evolved by exon shuffling of a family of human receptor domains. Nat Biotechnol 23, 1556–1561 (2005). https://doi.org/10.1038/nbt1166

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