Three parallel bioorthogonal reactions in one solution.a, b
a 50 μM 50, 50 μM 78, and 10 μM 82 in 5 mM phophate pH 7.5 with 25 mM NaCl at 37 °C in the presence of 50 mM each 3-methylbutyraldehyde and trans-cyclooctene and 40 mM cyclopentadiene. HPLC, top: starting material peptides; bottom: crude HPLC chromatogram after 30 minutes. All products were confirmed by isolation, mass spectrometry, and comparison to authentic material synthesized independently.
b RN = Ac-Thr-Tyr-, RC = -Asn-NH2.