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2 protocols using 2 boc amino ethanethiol

1

Immunoglobulin G Extraction and Proteomic Analysis

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Pentaerythritol-tetrakis(3-mercaptopropionate) (“tetrathiol”), triallyl-1,3,5-triazine-2,4,6-(1H,3H,5H)-trione (“triallyl”), 2-(boc amino) ethanethiol, and methanol (99.8%) were obtained from Sigma-Aldrich (Brøndby, Denmark). Lucirin TPO-L was obtained from BASF (Ludvigshafen, Germany). Acetonitrile (ACN), trifluoroacetic acid (TFA), DL-dithiothreitol (D9779, DTT), iodoacetamide (I670-9, IAA), immunoglobulin G (56,834-25MG, IgG, from human serum), trypsin (T1426, from bovine pancreas), and salts were purchased from Sigma-Aldrich (Stockholm, Sweden). L( +)-Ascorbic acid (99.0–100.5%) was from VWR chemicals (Stockholm, Sweden). Water (MQ H2O) was purified in a Millipore Synergy® 185 (Bedford, MA, USA) to a resistivity of 18.2 MΩ·cm at 25 °C. DEAE Affi-Gel® Blue Gel (DEAE) used for IgG extraction was obtained from Bio-RAD, Hercules, USA. Pierce® C18 Tips, 100 µL bed size, were from Thermo Fisher Scientific (Rockford, USA). Kinesis Tubing PEEK (TM) natural 1/32 inch × 0.015 inch connecting syringe and microchip was purchased from Fisher Scientific (Stockholm, Sweden). MALDI matrices 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxycinnamic acid (HCCA) and MALDI plate were obtained from Bruker Daltonics (Bremen, Germany).
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2

Synthesis of Functionalized Polymers

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Glycidyl methacrylate (GMA, 97%) and ethylene glycol dimethacrylate (EGDMA, 98%) were purchased from Sigma-Aldrich and passed through a column of neutral Al2O3 prior to use. N,N-dimethylformamide (DMF, anhydrous, 99.8%), trifluoroacetic acid (TFA, HPLC grade, >99.0%), N,N-dimethylethanolamine (redistilled, 99.5%), 1,4-dioxane (HPLC grade, >99.5%), azobisisobutyronitrile (AIBN, 98%), 2-(Boc-amino)ethanethiol (97%), lithium hydroxide (LiOH, reagent grade, 98%), tris((1-benzyl-4-triazolyl)methyl)amine (TBTA, 97%) and tetrakis(acetonitrile)copper(i) hexafluorophosphate (Cu(CH3CN)4PF6, 97%) were purchased from Sigma-Aldrich and used without further purification. Sodium azide (NaN3, 99%) was purchased from Acros Organics and used as supplied.

Safety note: Large scale reactions of azide are prone to dangerous exothermic reactions. The use of chlorinated solvents in the presence of sodium azide must be avoided, as the byproducts may be highly explosive.81

4-methyl-7-(prop-2-yn-1-yloxy)-2H-chromen-2-one was purchased from MolPort as used without any further purifications. Dichloromethane (DCM, 99.7%) and methanol (MeOH) were obtained from Ossum Chemicals. Demi-water was used unless stated otherwise. The RAFT agent 2-[[(butylthio)thioxomethyl] thio] propanoic acid was synthesized following a literature procedure.79
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