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Ramou, E, Rebordao G, Palma SICJ, Roque ACA.  2021.  Stable and Oriented Liquid Crystal Droplets Stabilized by Imidazolium Ionic Liquids. MOLECULES. 26(19):6044.PDF
Ramou, E, Palma SICJ, Roque ACA.  2022.  Nanoscale Events on Cyanobiphenyl-Based Self-Assembled Droplets Triggered by Gas Analytes. ACS Applied Materials and Interfaces. 14(4):6261-6273.PDF
Rebordão, G, Palma SICJ, Roque ACA.  2020.  Microfluidics in Gas Sensing and Artificial Olfaction. Sensors . 20(20):5742. AbstractPDF

Rapid, real-time, and non-invasive identification of volatile organic compounds (VOCs)
and gases is an increasingly relevant field, with applications in areas such as healthcare, agriculture,
or industry. Ideal characteristics of VOC and gas sensing devices used for artificial olfaction include
portability and affordability, low power consumption, fast response, high selectivity, and sensitivity.
Microfluidics meets all these requirements and allows for in situ operation and small sample amounts,
providing many advantages compared to conventional methods using sophisticated apparatus such
as gas chromatography and mass spectrometry. This review covers the work accomplished so far
regarding microfluidic devices for gas sensing and artificial olfaction. Systems utilizing electrical
and optical transduction, as well as several system designs engineered throughout the years are
summarized, and future perspectives in the field are discussed.

Rodrigues, R, Palma SICJ, Correia VJ, Padrao I, Pais J, Banza M, Alves C, Deuermeier J, Martins C, Costa HMA, Ramou E, Silva Pereira C, Roque ACA.  2020.  Sustainable plant polyesters as substrates for optical gas sensors. Materials Today Bio. 8:100083. AbstractPDF

The fast and non-invasive detection of odors and volatile organic compounds (VOCs) by gas sensors and electronic
noses is a growing field of interest, mostly due to a large scope of potential applications. Additional drivers for the
expansion of the field include the development of alternative and sustainable sensing materials. The discovery
that isolated cross-linked polymeric structures of suberin spontaneously self-assemble as a film inspired us to
develop new sensing composite materials consisting of suberin and a liquid crystal (LC). Due to their stimuliresponsive and optically active nature, liquid crystals are interesting probes in gas sensing. Herein, we report
the isolation and the chemical characterization of two suberin types (from cork and from potato peels) resorting to
analyses of gas chromatography–mass spectrometry (GC-MS), solution nuclear magnetic resonance (NMR), and Xray photoelectron spectroscopy (XPS). The collected data highlighted their compositional and structural differences. Cork suberin showed a higher proportion of longer aliphatic constituents and is more esterified than potato
suberin. Accordingly, when casted it formed films with larger surface irregularities and a higher C/O ratio. When
either type of suberin was combined with the liquid crystal 5CB, the ensuing hybrid materials showed distinctive
morphological and sensing properties towards a set of 12 VOCs (comprising heptane, hexane, chloroform,
toluene, dichlormethane, diethylether, ethyl acetate, acetonitrile, acetone, ethanol, methanol, and acetic acid).
The optical responses generated by the materials are reversible and reproducible, showing stability for 3 weeks.
The individual VOC-sensing responses of the two hybrid materials are discussed taking as basis the chemistry of
each suberin type. A support vector machines (SVM) algorithm based on the features of the optical responses was
implemented to assess the VOC identification ability of the materials, revealing that the two distinct suberin-based
sensors complement each other, since they selectively identify distinct VOCs or VOC groups. It is expected that
such new environmentally-friendly gas sensing materials derived from natural diversity can be combined in arrays
to enlarge selectivity and sensing capacity.

Roque, ACA, Fred A, Gamboa H.  2019.  Foreword, January 2019. BIODEVICES 2019 - 12th International Conference on Biomedical Electronics and Devices, Proceedings; Part of 12th International Joint Conference on Biomedical Engineering Systems and Technologies, BIOSTEC 2019. , Prague: SciTePress
Roque, ACA, Lowe CR.  2007.  Rationally designed ligands for use in Affinity Chromatography: An artificial Protein L. Affinity Chromatography: Methods and Protocols. (M. Zachariou, Ed.).:93-110., U.S.A.: Humana Press Inc. Abstract

Synthetic affinity ligands can circumvent the drawbacks of natural immunoglobulin (Ig)-binding proteins by imparting resistance to chemical and biochemical degradation and to in situ sterilization, as well as ease and low cost of production. Protein L (PpL), isolated from Peptostreptococcus magnus strains, interacts with the Fab (antigen-binding fragment) portion of Igs, specifically with kappa light chains, and represents an almost universal ligand for the purification of antibodies. The concepts of rational design and solid-phase combinatorial chemistry were used for the discovery of a synthetic PpL mimic affinity ligand. The procedure presented in this chapter represents a general approach with the potential to be applied to different systems and target proteins.

Roque, ACA, Wilson OC.  2008.  Adsorption of gum Arabic on bioceramic nanoparticles. Materials Science & Engineering C.- Biomimetic and Supramolecular Systems. 28:443–447., Number 3 Abstract


Roque, ACA, Bicho A, Batalha IL, Cardoso AS, Hussain A.  2009.  Biocompatible and bioactive gum Arabic coated iron oxide magnetic nanoparticles. Journal of Biotechnology. 144:313–320., Number 4 AbstractWebsite

The surface modification of iron oxide magnetic nanoparticles {(MNPs)} with gum Arabic {(GA)} via adsorption and covalent coupling was studied. The adsorption of {GA} was assessed during {MNP} chemical synthesis by the co-precipitation method {(MNP\_GA)}, and after {MNP} synthesis on both bare magnetite and {MNP\_GA.} The covalent immobilization of {GA} at the surface of aldehyde-activated {(MNP\_GAAPTES)} or aminated {MNPs} {(MNP\_GAEDC)} was achieved through free terminal amino and carboxylate groups from {GA.} The presence of {GA} at the surface of the {MNPs} was confirmed by {FTIR} and by the quantification of {GA} by the bicinchoninic acid test. Results indicated that the maximum of {GA} coating was obtained for the covalent coupling of {GA} through its free carboxylate groups {(MNP\_GAEDC)}, yielding a maximum of 1.8&\#xa0;g of {GA} bound/g of dried particles. The hydrodynamic diameter of {MNPs} modified with {GA} after synthesis resulted in the lowest values, in opposition to the {MNPs} co-precipitated with {GA} which presented the tendency to form larger aggregates of up to 1&\#xa0;μm. The zeta potentials indicate the existence of negatively charged surfaces before and after {GA} coating. The potential of the {GA} coated {MNPs} for further biomolecule attachment was assessed through anchorage of a model antibody to aldehyde-functionalized {MNP\_GA} and its subsequent detection with an {FITC} labeled anti-antibody.

Roque, ACA, Pina AS, Azevedo AM, Aires-Barros R, Jungbauer A, Profio DG, Heng JYY, Haigh J, Ottens M.  2020.  Anything but Conventional Chromatography Approaches in Bioseparation. Biotechnology Journal. (e1900274):1-8.
Roque, ACA.  2009.  Ligand-Macromolecule Interactions in Drug Discovery. , U.S.A.: Methods in Molecular Biology, Humana Press Inc.Website
Roque, ACA, Bispo S, Pinheiro ARN, Antunes JMA, Gonçalves D, Ferreira HA.  2009.  Antibody immobilization on magnetic particles. Journal of Molecular Recognition. 22:77–82., Number 2 AbstractWebsite

Magnetic particles {(MNPs)} offer attractive possibilities in biotechnology. {MNPs} can get close to a target biological entity, as their controllable sizes range from a few nanometres up to tens of nanometres, and their surface can be modified to add affinity and specificity towards desired molecules. Additionally, they can be manipulated by an external magnetic field gradient. In this work, the study of ferric oxide {(Fe3O4)} {MNPs} with different coating agents was conducted, particularly in terms of strategies for antibody attachment at the surfaces (covalent and physical adsorption) and the effects of blocking buffer composition and incubation times on the specific and non-specific interactions observed. The considered biological model system consisted of a coating antibody (goat {IgG)}, bovine serum albumin {(BSA)} as blocking agent, and a complementary antibody labelled with {FITC} (anti-goat {IgG).} The detection of antibody binding was followed by fluorescence microscopy and the intensity of the signals quantified. The ratio between the mean grey values of negative and positive controls, as well as the maximum intensity attainable in positive controls, were considered in the evaluation of the assays efficiency. The covalent immobilization of the coating antibody was more successful as opposed to protein adsorption. For covalent immobilization, silica-coated {MNPs}, a 5% (w/v) concentration of {BSA} in the blocking buffer and incubation times of 1 h produced the best results in terms of assay sensitivity. However, when conducting the assay for incubation periods of 10 min, the fluorescence signal was reduced by 44% but the assay specificity was maintained.

Roque, ACA, Lowe CR.  2007.  Affinity chromatography: History, Perspectives, Limitations and Prospects. Affinity Chromatography: Methods and Protocols. (M. Zachariou, Ed.).:1-23., U.S.A.: Humana Press Inc. Abstract

Biomolecule separation and purification has until very recently steadfastly remained one of the more empirical aspects of modern biotechnology. Affinity chromatography, one of several types of adsorption chromatography, is particularly suited for the efficient isolation of biomolecules. This technique relies on the adsorbent bed material that has biological affinity for the substance to be isolated. This review is intended to place affinity chromatography in historical perspective and describe the current status, limitations and future prospects for the technique in modern biotechnology.

Roque, ACA, Silva CSO, Taipa ÂM.  2007.  Affinity-based methodologies and ligands for antibody purification: Advances and perspectives. Journal of Chromatography A. 1160:44–55., Number 1-2 AbstractWebsite

Many successful, recent therapies for life-threatening diseases such as cancer and rheumatoid arthritis are based on the recognition between native or genetically engineered antibodies and cell-surface receptors. Although naturally produced by the immune system, the need for antibodies with unique specificities and designed for single application, has encouraged the search for novel antibody purification strategies. The availability of these products to the end-consumer is strictly related to manufacture costs, particularly those attributed to downstream processing. Over the last decades, academia and industry have developed different types of interactions and separation techniques for antibody purification, affinity-based strategies being the most common and efficient methodologies. The affinity ligands utilized range from biological to synthetic designed molecules with enhanced resistance and stability. Despite the successes achieved, the purification “paradigm” still moves interests and efforts in the continuous demand for improved separation performances. This review will focus on recent advances and perspectives in antibody purification by affinity interactions using different techniques, with particular emphasis on affinity chromatography.