Please cite this article in press as: M.A. Mata-Gómez, et al., Synthesis of adsorbents with dendronic structures for protein hydrophobic interaction chromatography, J. Chromatogr. A (2016), http://dx.doi.org/10.1016/j.chroma.2016.03.057 ARTICLE IN PRESS G Model CHROMA-357414; No. of Pages 10 Journal of Chromatography A, xxx (2016) xxx–xxx Contents lists available at ScienceDirect Journal of Chromatography A j o ur na l ho me page: www.elsevier.com/locate/chroma Synthesis of adsorbents with dendronic structures for protein hydrophobic interaction chromatography Marco A. Mata-Gómez a , Sena Yaman b , Jesus A. Valencia-Gallegos a , Canan Tari c , Marco Rito-Palomares a,∗ , José González-Valdez a,∗ a School of Engineering and Sciences, Tecnologico de Monterrey, Ave. Eugenio Garza Sada 2501 Sur, Monterrey, NL 64849, Mexico b Izmir Institute of Technology, Department of Bioengineering, Urla, 35430 ˙ Izmir, Turkey c Izmir Institute of Technology, Department of Food Engineering, Urla, 35430 ˙ Izmir, Turkey a r t i c l e i n f o Article history: Received 2 February 2016 Received in revised form 16 March 2016 Accepted 20 March 2016 Available online xxx Keywords: Dendrons Hydrophobic interaction chromatography Adsorbent Resin a b s t r a c t 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 per- formance 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 cou- pled 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 mol ligand/mL resin for RG3 and RG5, respec- tively. Static adsorption capacity of modified resins was found to be ∼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, -lactoglobulin and -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 chromato- graphic 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. © 2016 Elsevier B.V. All rights reserved. 1. Introduction A major need in the biotech industry is the isolation and purifica- tion of proteins from complex mixtures where protein downstream processing may account for up to an 80% of the final product total cost. Hence, the development of novel isolation and purification methodologies or the optimization of the current standardized protocols to achieve the highest purities and yields, raises mayor attention in the design of bioprocesses. In this context, chromato- graphic methods are the most used techniques for purification of proteins due to the high recovery and purities achieved. Among ∗ Corresponding authors. E-mail addresses: matago24@gmail.com (M.A. Mata-Gómez), senaymn@gmail.com (S. Yaman), valencia@itesm.mx (J.A. Valencia-Gallegos), canantari@iyte.edu.tr (C. Tari), mrito@itesm.mx (M. Rito-Palomares), jose gonzalez@itesm.mx (J. González-Valdez). these, size exclusion chromatography (SEC), hydrophobic interac- tion chromatography (HIC), ion-exchange chromatography (IEX), reverse phase chromatography (RPC) and affinity chromatography (AC) are the most common used protein separation chromato- graphic techniques [1,2]. HIC is a powerful and widely used technique for purification of proteins [3,4]. It is a key methodology when purification of mon- oclonal antibodies [5–7] is required. It is often used in the final polishing step of downstream processes as it can easily remove high-molecular weight aggregates [8]. HIC relies on the inter- action between the hydrophobic regions on the surface of the biomolecules and the ligands on the support under high concentra- tion of salts i.e. ammonium sulfate or sodium chloride [3]. There are numerous commercially available adsorbents with different types of hydrophobic ligands i.e. butyl, octyl and phenyl groups. How- ever, one of the major drawbacks in HIC is the low ligand density that impacts directly on the adsorption capacity of resins. Further- more, the methods used to activate the support materials might be http://dx.doi.org/10.1016/j.chroma.2016.03.057 0021-9673/© 2016 Elsevier B.V. All rights reserved.