PdBT was click-conjugated by alkyne-azide cycloaddition to CS, BMPm, or NC by mixing the biomolecule with PdBT in water at ambient temperature (Fig. 2B). Briefly, the biomolecule of interest was mixed with 0.5 mol eq. of PdBT, 2 mol eq. of DTT for disulfide inhibition, and 0.1 mol eq. of Cp*Ru(cod)Cl in ultrapure H2O, followed by stirring at ambient temperature for 8 hours. Following this, unreacted biomolecules, unmodified PdBT, and impurities were removed by dialysis for 24 hours at either 2-kDa MWCO for BMPm/PdBT and NC/PdBT or 50-kDa MWCO for CS/PdBT, with replacement of H2O every 6 to 8 hours. After dialysis, solutions were flash-frozen in liquid N2 and lyophilized.

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