Colloids and Surfaces B: Biointerfaces 161 (2018) 645–653 Contents lists available at ScienceDirect Colloids and Surfaces B: Biointerfaces j o ur nal ho me pa ge: www.elsevier.com/locate/colsurfb Full Length Article OBP fused with cell-penetrating peptides promotes liposomal transduction Filipa Gonc ¸ alves a , Tarsila G. Castro a , Eugénia Nogueira a , Ricardo Pires b,c , Carla Silva a , Artur Ribeiro a , Artur Cavaco-Paulo a,∗ a Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal b 3B´ ıs Research Group, Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Ave Park, 4806-909 Taipas, Guimarães, Portugal c ICVS/3B’s, PT Government Associate Laboratory, Braga, Guimarães, Portugal a r t i c l e i n f o Article history: Received 22 May 2017 Received in revised form 2 November 2017 Accepted 8 November 2017 Available online 10 November 2017 Keywords: 1-aminoanthracene (1-AMA) Cell-penetrating peptides Odorant-binding protein 1-AMA transduction Liposomes a b s t r a c t Cell-penetrating peptides (CPPs) have been applied as novel transport systems with the ability to facil- itate the delivery of peptides, proteins, and oligonucleotides into cells. Herein, we designed different fusion proteins composed by pig odorant binding protein (OBP-I) and three CPPs, namely Tat, pVEC and Pep-1. A new methodology using liposomes as reservoirs and OBP:CPPs as carriers was developed as an advanced system to capture odorant molecules. 1-aminoanthracene (1-AMA) was used as a model molecule to evaluate the transduction ability of OBP:CPPs into the reservoirs. The transduction efficiency was dependent on the initial capacity of OBP:CPPs to bind 1-AMA and on the penetration of liposomes promoted by the CPPs. An encapsulation efficiency of 42% was obtained with OBP:Tat fusion protein. The presence of Tat peptide increased the 1-AMA transduction of 1.3 and 2.5 fold compared with Pep-1 and pVEC, respectively. This work expands the application of OBPs and CPPs on the design of promising capture and delivery systems for textile and cosmetic applications. © 2017 Elsevier B.V. All rights reserved. 1. Introduction CPPs are cationic peptides, normally up to 30 residues, which can be amphipathic or hydrophobic, possessing low cellular toxicity [1]. These peptides are widely studied to deliver biologically active molecules into cells, such as peptides, proteins, RNA, DNA, oligonu- cleotides and liposomes without the need of specific membrane receptors [1–3]. Several works report the use of CPPs in biomedical applications. Tat-conjugated quantum dots administrated intra- arterially at a proximal cervical carotid artery in rats were able to cross the highly impermeable blood-brain barrier [4]. Elmquist et al. (2001) demonstrated the internalization of pVEC labeled with fluorescein isothiocyanate (FITC) into three different endothelial cell lines after treatment with this peptide [5]. Jing et al. (2016) demonstrated that the combined use of CPPs-loaded nanobub- bles with ultrasound-targeted microbubble destruction (UTMD) technology could efficiently improve gene transfection in cultured breast cancer TNBC cells [6]. The application of CPPs in other areas rather than therapeutics, such as textile functionalization, was ∗ Corresponding author. E-mail address: artur@deb.uminho.pt (A. Cavaco-Paulo). exploited herein for the first time. CPPs were conjugated with an odorant-binding protein (OBP) for the capture and transduction of odorant molecules. Liposomes were used as final reservoirs and 1-AMA as the model molecule. Odorant-binding protein I (PDB 1DZK code) is a transport pro- tein present in the nasal mucosa of pig constituted by 157 amino acids. This protein was selected for this study based on the infor- mation available about the three-dimensional structure and the binding specificity for a large number of natural and synthetic molecules [7,8]. OBP-I is composed by eight antiparallel -sheets [9], forming an internal cavity to bind different ligands, like ter- penoids, aromatic compounds, aliphatic molecules and aldehydes [7]. OBPs have been studied in several applications. Wei et al. (2008) designed different mutants of pig OBP to bind several aromatic polycyclic hydrocarbons. This study opened the view for the use of OBPs as biosensors for the monitoring of aromatic pollutants [10]. Di Pietrantonio et al. (2013) used surface acoustic wave (SAW) biosensor systems with three different OBPs as probes for the detec- tion of low concentration of octanol (13 ppm) and carvone (9 ppm) [11]. More recently, Silva et al. (2014) used pig OBP for the reduc- tion of unpleasant odors and controlled release of fragrances when immobilized in fabric supports [12]. https://doi.org/10.1016/j.colsurfb.2017.11.026 0927-7765/© 2017 Elsevier B.V. All rights reserved.