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Rapid and low-cost multiplex synthesis of chemokine analogs

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Published in Journal of Biological Chemistry. 2018, vol. 293, no. 49, p. 19092-19100
Abstract Peptides represent a promising source of new medicines, but improved technologies are needed to facilitate discovery and optimization campaigns. In particular, longer peptides with multiple disulfide bridges are challenging to produce, and producing large numbers of structurally related variants is dissuasively costly and time consuming. The principal cost and time drivers are the multiple column chromatography purification steps that are used during the multi-step chemical synthesis procedure, which involves both ligation and oxidative refolding steps. In this study we developed a method for multiplex parallel synthesis of complex peptide analogs in which the structurally variant region of the molecule is produced as a small peptide on a 384-well synthesizer, with subsequent ligation to the longer, structurally invariant region and oxidative refolding carried out in-well without any column purification steps. To test the method we used a panel of 96 analogs of the chemokine RANTES/CCL5 (69 residues, two disulfide bridges) were synthesized using standard approaches and characterized pharmacologically in an earlier study. Although, as expected, the multiplex method generated chemokine analogs of lower purity than those produced in the original study, it was nonetheless possible to closely match the pharmacological attributes (anti-HIV potency, capacity to elicit G protein signaling, capacity to elicit intracellular receptor sequestration) of each chemokine analog to reference data from the earlier study. This rapid, low-cost approach has the potential to support discovery and optimization campaigns based on analogs of other chemokines, as well as those of other complex peptide and small protein targets of a similar size.
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PMID: 30305389
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Research group Hiv (835)
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PAOLINI-BERTRAND, Marianne et al. Rapid and low-cost multiplex synthesis of chemokine analogs. In: Journal of Biological Chemistry, 2018, vol. 293, n° 49, p. 19092-19100. https://archive-ouverte.unige.ch/unige:115805

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Deposited on : 2019-04-08

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