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Both previous examples had possessed alanine at the position for backbone protection.
The site of backbone protection is no longer restricted to only two residues.
The results demonstrate successful incorporation of backbone protection and improved synthesis (ESI, † Fig. S2).
The synthesis of peptides rich in aggregation prone sequences can be improved with backbone protection.
For comparison, the test sequence was synthesised without backbone protection on rink amide resin (Fig. 1A).
We report the automated introduction of backbone protection to a peptide.
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Backbone amide protection was first investigated (Dmb 2 Scheme 1) for its dramatic effect on peptide solubility.
Application of the new methodology to the top-down HDX MS characterization of a small (99 residue long) disulfide-containing protein β2-microglobulin allows the backbone amide protection to be probed with nearly a single-residue resolution.
Strategies to combat Fmoc-SPPS resin-bound aggregation and improve synthetic product yield include the use of more potent acylation reagents, 25 backbone amide protection, 26, 27 incorporation of pseudoproline, 28, 29 microwave irradiation to enhance coupling efficiency, 30 SPPS at elevated temperatures, 30 and use of DBU instead of piperidine to deprotect the N-terminal Fmoc group.
Application of the new methodology to the top-down HDX MS characterization of a small (99 residue long) disulfide-containing protein β2-microglobulin allowed the backbone amide protection to be probed with nearly a single-residue resolution across the entire sequence.
These algorithms are the backbone of data protection and secrecy for highly sensitive and classified data.
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