Synthesis of adsorbents with dendronic structures for protein hydrophobic interaction chromatography

Here, we introduced a new technology based on the incorporation of dendrons-branched chemical structures-onto supports for synthesis of HIC adsorbents. In doing so we studied the synthesis and performance of these novel HIC dendron-based adsorbents. The adsorbents were synthesized in a facile two-step reaction. First, Sepharose 4FF (R) was chemically modified with polyester dendrons of different branching degrees i.e. third (G3) or fifth (G5) generations. Then, butyl-end valeric acid ligands were coupled to dendrons via ester bond formation. UV-vis spectrophotometry and FTIR analyses of the modified resins confirmed the presence of the dendrons and their ligands on them. Inclusion of dendrons allowed the increment of ligand density, 82.5 +/- 11 and 175.6 +/- 5.7 mu mol ligand/mL resin for RG3 and RG5, respectively. Static adsorption capacity of modified resins was found to be similar to 60 mg BSA/mL resin. Interestingly, dynamic binding capacity was higher at high flow rates, 62.5 +/- 0.8 and 58.0 +/- 0.5 mg/mL for RG3 and RG5, respectively. RG3 was able to separate lipase, beta-lactoglobulin and a-chymotrypsin selectively as well as fractionating of a whole proteome from yeast. This innovative technology will improve the existing HIC resin synthesis methods. It will also allow the reduction of the amount of adsorbent used in a chromatographic procedure and thus permit the use of smaller columns resulting in faster processes. Furthermore, this method could potentially be considered as a green technology since both, dendrons and ligands, are formed by ester bonds that are more biodegradable allowing the disposal of used resin waste in a more ecofriendly manner when compared to other exiting resins. (C) 2016 Elsevier B.V. All rights reserved.