A platform for efficient, thiol-stable conjugation to albumin's native single accessible cysteine† †Electronic supplementary information (ESI) available: LC-MS, ES-MS and deconvoluted spectra for all reactions with proteins described herein, and 1H and 13C NMR spectra for all small molecule constructs. See DOI: 10.1039/c5ob01205h Click here for additional data file.
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چکیده
All reagents were purchased from Sigma-Aldrich or Life Technologies and were used as received. Where described below pet. refers to petroleum ether (40–60 °C). All reactions were monitored by thin-layer chromatography (TLC) on pre-coated SIL G/UV254 silica gel plates (254 m) purchased from VWR. Flash column chromatography was carried out with Kiesegel 60 M, 0.04/0.063 mm (200–400 mesh) silica gel. 1 H and 13 C NMR spectra were recorded at ambient temperature on a Bruker Avance 600 instrument operating at a frequency of 600 MHz for 1 H and 150 MHz for 13 C in CDCl3. The chemical shifts () for 1 H and 13 C are quoted relative to residual signals of the solvent on the ppm scale. 1 H NMR peaks are reported as singlet (s), doublet (d), triplet (t), quartet (q), quintet (quint.), sextet (sext.), octet (oct.), multiplet (m), broad (br), doublet of doublets (dd), doublet of triplets (dt), AB quartet (ABq). Coupling constants (J values) are reported in Hertz (Hz) and are H-H coupling constants unless otherwise stated. Signal multiplicities in 13 C NMR were determined using the distortionless enhancement by phase transfer (DEPT) spectral editing technique. Infrared spectra were obtained on a Perkin Elmer Spectrum 100 FTIR Spectrometer operating in ATR mode with frequencies given in reciprocal centimetres (cm -1 ). Melting points were measured with a Gallenkamp apparatus and are uncorrected. Mass spectra were obtained on a VG70-SE mass spectrometer. Recombinant native sequence albumin 1 was obtained from Novozymes Biopharma.
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