Peptide purity was determined as described previously [29 (link)] by analytical RP-HPLC using a Waters 717 plus Autosampler, In-line Degasser AF, 600 Controller, and 2996 Photodiode Array Detector; the column used was a Waters Symmetry C18 column (5 μm, 4.6 mm × 250 mm, Milford, MA, USA). An aqueous acetonitrile (ACN) gradient with 0.1% TFA added (mixing eluent A: H2O + 0.1% TFA and eluent B: 90:10 ACN-H2O + 0.1% TFA) was employed using milli-Q water. Preparative RP-HPLC was performed by using a Waters XSelect Peptide CSH C18 OBDTM column (5 μm, 19 mm × 250 mm, Milford, MA, USA) with the same eluents as for analytical HPLC.
MicroflexTM (Bruker Corporation, Bremen, Germany) equipped with FlexControl software (Bruker Daltonik GmbH, Bremen, Germany) was used to obtain the MALDI-TOF-MS spectra, and the data were processed using flexAnalysis (Bruker Daltonik GmbH). All of the reagents and solvents were used without further purification.
MicroflexTM (Bruker Corporation, Bremen, Germany) equipped with FlexControl software (Bruker Daltonik GmbH, Bremen, Germany) was used to obtain the MALDI-TOF-MS spectra, and the data were processed using flexAnalysis (Bruker Daltonik GmbH). All of the reagents and solvents were used without further purification.
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