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Synthesis of 99 aa peptide P3‐P1‐P3 (41) by ligation of 54 aa thioester 38 and 55 aa MPyE‐peptide 39 and auxiliary removal. UPLC analysis of aliquots withdrawn A) at the start of the ligation, B) after 24 h ligation and treatment with 2.5 % hydrazine (to confirm completion of S→N acyl transfer), C) auxiliary removal from purified ligation product and D) of purified 41.
