2 2 aminoethoxy ethanol

For synthesis of KAM-derivatized resin, packed NHS-activated sepharose 4 fast flow resin (volume = 2 mL; GE Healthcare) was washed with anhydrous DMSO (3 × 10 mL). To the washed NHS-activated sepharose resin was added 0.5 mM KAM in anhydrous DMSO (8 mL; 2 μmol compound/mL of resin), followed by the addition of triethylamine (30 μL). The reaction mixture was vortexed to mix and pelleted by centrifugation (100 x g, 2 min). An aliquot of the supernatant (50 μL) was saved for LC/MS analysis. The reaction mixture was allowed incubate overnight at room temp with end-over-end rotating agitation. On the following day, the reaction mixture was pelleted by centrifugation (100 x g, 2 min). An aliquot of the supernatant (50 μL) was saved for LC/MS analysis. Completion of coupling was inferred by loss of starting material following LC/MS analysis. 2-(2-Aminoethoxy)ethanol (100 μL; Sigma-Aldrich) was added to the reaction mixture, vortexed, and incubate overnight at room temp with end-over-end agitation. The KAM-derivatized resin was then washed with anhydrous DMSO (3 × 10 mL) and 95% EtOH (3 × 10 mL).
For synthesis of imatinib-derivatized resin, a similar protocol was followed as described above except that final concentration of compound on the bead was 0.25 μmol compound/mL.
For synthesis of dasatinib-derivatized resin, the protocol for the KAM-derivatized resin was followed.

Horseradish peroxidase (HRP) (EC 1.11.1.7, from Armoracia rusticana, 500 U·mg⁻¹), tetrachloroauric acid trihydrate, sodium salt of ABTS, sodium citrate, N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), pentafluorophenol (PFP), 2-(2-aminoethoxy) ethanol (AEE), dimethylformamide (DMF), N,N-diisopropylethylamine (DIPEA), 16-mercaptohexadecanoic acid (MHDA), methylamine (MA), dimethylamine (DMA), trimethylamine (TMA), NaCl, KH₂PO₄, Na₂HPO₄, chloroform, and Butvar solution B-98 were purchased from Sigma-Aldrich (ALSI Ltd., Kiev, Ukraine). The cathodic electrodeposition paint “GY 83–0270 0005” was from BASF Farben und Lacke (Munster, Germany). All buffers and standard solutions were prepared using the water purified by the Milli-Q system (Millipore).

FeCl₂·4H₂O, FeCl₃·6H₂O, malonic acid, 2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HATU), N,N-diisopropylethylamine (DIEA), and dimethylformamide (DMF) were from Bomaijie Inc. (Beijing, China). Glucose, glutamine, ammonium hydroxide (28~30 wt%), tetraethylorthosilicate (TEOS), (3-aminopropyl) triethoxysilane (APS), 2-(2-aminoethoxy) ethanol, 4-(trans-2-carboxyvinyl) phenylboronic acid, and ovalbumin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Water was purified on a Milli-Q system (Millipore, Milford, MA, USA). PNGase F was purchased from New England Lab. Human milk samples were provided by a healthy mother, attending the First Affiliated Hospital of Medical College, Peking University, China.