A fully automated flow-based approach for accelerated peptide synthesis.

Here we report a fully automated, flow-based approach to solid-phase polypeptide synthesis, with amide bond formation in 7 seconds and total synthesis times of 40 seconds per amino acid residue. Crude peptide purities and isolated yields were comparable to those for standard-batch solid-phase peptide synthesis. At full capacity, this approach can yield tens of thousands of individual 30-mer peptides per year.

[1]  R. Sheppard,et al.  A reversible protecting group for the amide bond in peptides. Use in the synthesis of ‘difficult sequences’ , 1993 .

[2]  E. Kaiser,et al.  Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. , 1970, Analytical biochemistry.

[3]  P. Alewood,et al.  Evaluation of COMU as a coupling reagent for in situ neutralization Boc solid phase peptide synthesis , 2012, Journal of peptide science : an official publication of the European Peptide Society.

[4]  Yongxin Han,et al.  Occurrence and Minimization of Cysteine Racemization during Stepwise Solid-Phase Peptide Synthesis(1)(,)(2). , 1997, The Journal of organic chemistry.

[5]  R. B. Merrifield,et al.  Quantitative monitoring of solid-phase peptide synthesis by the ninhydrin reaction. , 1981, Analytical biochemistry.

[6]  H. Fabian,et al.  Synthesis of ‘difficult’ peptide sequences: application of a depsipeptide technique to the Jung–Redemann 10- and 26-mers and the amyloid peptide Aβ(1–42) , 2004 .

[7]  P. White,et al.  An accurate method for the quantitation of Fmoc-derivatized solid phase supports , 2002, Letters in peptide science.

[8]  J. Meienhofer,et al.  Special methods in peptide synthesis , 1980 .

[9]  M. Mutter,et al.  Pseudo-prolines in peptide synthesis: Direct insertion of serine and threonine derived oxazolidines in dipeptides , 1995 .

[10]  Shui-Tein Chen,et al.  Enhanced coupling efficiency in solid-phase peptide synthesis by microwave irradiation , 1992 .

[11]  W. Chan,et al.  Fmoc Solid Phase Peptide Synthesis: A Practical Approach (Practical Approach Series) , 2019 .

[12]  Steven V. Ley,et al.  ReactIR Flow Cell: A New Analytical Tool for Continuous Flow Chemical Processing , 2010 .

[13]  Steffen Hardt,et al.  Helical Flows and Chaotic Mixing in Curved Micro Channels , 2004 .

[14]  M. Beyermann,et al.  Solid-phase peptide synthesis: from standard procedures to the synthesis of difficult sequences , 2007, Nature Protocols.

[15]  E. Bayer,et al.  Polystyrene-Immobilized PEG Chains , 1992 .

[16]  Samuel Zalipsky,et al.  Poly(ethylene glycol): Chemistry and Biological Applications , 1997 .

[17]  B. Pentelute,et al.  Rapid Total Synthesis of DARPin pE59 and Barnase , 2014, Chembiochem : a European journal of chemical biology.

[18]  R. Sheppard,et al.  Peptide synthesis. Part 11. A system for continuous flow solid phase peptide synthesis using fluorenylmethoxycarbonyl-amino acid pentafluorophenyl esters , 1988 .

[19]  Alexander M. Spokoyny,et al.  Rapid Flow‐Based Peptide Synthesis , 2014, Chembiochem : a European journal of chemical biology.

[20]  S. Y. Wong,et al.  On-demand continuous-flow production of pharmaceuticals in a compact, reconfigurable system , 2016, Science.

[21]  Chemical synthesis of peptides and proteins. , 1988, Annual review of biochemistry.

[22]  Jun-ichi Yoshida,et al.  Flash chemistry: flow chemistry that cannot be done in batch. , 2013, Chemical communications.

[23]  Jonathan M. Collins,et al.  High-efficiency solid phase peptide synthesis (HE-SPPS). , 2014, Organic letters.

[24]  Peter H Seeberger,et al.  Applying flow chemistry: methods, materials, and multistep synthesis. , 2013, The Journal of organic chemistry.

[25]  C. Oliver Kappe,et al.  Continuous‐Flow Microreactor Chemistry under High‐Temperature/Pressure Conditions , 2009 .

[26]  S. Kent,et al.  Enhanced Solvation of Peptides Attached to "Solid-Phase" Resins: Straightforward Syntheses of the Elastin Sequence Pro-Gly-Val-Gly-Val-Pro-Gly-Val-Gly-Val. , 2015, Organic letters.

[27]  M. T. Miranda,et al.  Solid-phase peptide synthesis at elevated temperatures: a search for and optimized synthesis condition of unsulfated cholecystokinin-12. , 2009, The journal of peptide research : official journal of the American Peptide Society.

[28]  Stephen B. H. Kent,et al.  In situ neutralization in Boc-chemistry solid phase peptide synthesis. Rapid, high yield assembly of difficult sequences. , 2009 .