Biomolecular Engineering Lab

Conference Affinity 2013

Prof. Cecília Roque, Ricardo Branco, Íris Batalha and Ana Margarida Dias participated in the conference Affinity 2013 - 20th Biennial Meeting of the International Society for Molecular Recognition held in Vienna (Austria) from the 26th to the 29th of June.

 

ESF-EMBO Symposium -“Molecular Perspectives on PPI"

From 25th to 30th May 2013, Ana Margarida Dias was present in the ESF-EMBO Symposium – “Molecular Perspectives on Protein-Protein Interactions” in Pultusk, Poland.

Funding and Facilities

Funding

The Biomolecular Engineering Group is funded by Fundação para a Ciência e a Tecnologia (Portugal) in 0.4 M€ and by awards granted by private and public entities (35 000 €).

Congratulations Dr. Ana Pina!

Our colleague Ana Pina is already a Dr.! 

She passed her Viva Voice Examination last 22nd May. We are very happy for her and wish her all the best!

2013 Annual Group Day

This year we had our "Open Day" in Serra da Arrábida, Setúbal region.

We went for an entire morning of cannoying. After all this physical activity we had lunch in the beach and enjoyed the sun.

Our group was in ECCE9-ECAB2 2013 at Den Haag (the Netherlands)

Last 21-25 April, our Group was at the 9th European Congress of Chemical Engineering, in the the Netherlands  (http://www.ecce2013.eu/).

Prof. Cecília Roque presented an Oral Communication, entitled "Toward a Sustainable Alternative for Antibody Purification: Theoretical and Experimental Studies", a result from the work of the authours A.C.A. Roque, A. Aguiar-Ricardo, A.M.G.C. Dias, I.L.Batalha, R.J.F.Branco, S.D.S.Santana, T. Barroso and V.L.Dhadge.

Expo FCT 2013

Like in the previous years, our group participated in Expo FCT.

Every year, this event brings thousands of high school students to the campus to get to know the curricular opportunies and research activities available at FCT.

Our group at....

Branco, R. J. F., A. M. G. C. Dias and A. C. A. Roque “Understanding the molecular recognition between antibody fragments and protein A biomimetic ligand” EMBO Conference - Catalytic Mechanisms by Biological Systems, 7-10 October, 2012, Groningen, The Netherlands.

Our participation at the ESBES 2012 conference

The group was at the ESBES + ISPPP 2012 Symposia 2012 in Istanbul/Turkey (23rd-26th September 2012) http://events.dechema.de/events/en/BEST2012.html.

Telma Barroso presented a keynote lecture:

T. Barroso, A.C.A. Roque, A. Aguiar-Ricardo, “Bioinspired affinity monoliths: a fast and efficient alternative system for antibody purification”

Margarida Dias presented an oral communication:

Santana, SDF, Dhadge VL, Roque ACA.  2012.  Dextran-Coated Magnetic Supports Modified with a Biomimetic Ligand for IgG Purification. ACS Applied Materials and Interfaces. 4(11):5907–5914. AbstractWebsite

extran-coated iron oxide magnetic particles modified with ligand 22/8, a protein A mimetic ligand, were prepared and assessed for IgG purification. Dextran was chosen as the agent to modify the surface of magnetic particles by presenting a negligible level of nonspecific adsorption. For the functionalization of the particles with the affinity ligand toward antibodies, three methods have been explored. The optimum coupling method yielded a theoretical maximum capacity for human IgG calculated as 568 ± 33 mg/g and a binding affinity constant of 7.7 × 104 M–1. Regeneration, recycle and reuse of particles was also highly successful for five cycles with minor loss of capacity. Moreover, this support presented specificity and effectiveness for IgG adsorption and elution at pH 11 directly from crude extracts with a final purity of 95% in the eluted fraction.

Sandu, ICA, Schäfer S, Magrini D, Bracci S, Roque ACA.  2012.  Cross-Section and Staining-Based Techniques for Investigating Organic Materials in Painted and Polychrome Works of Art: A Review.. Microscopy and Microanalysis. 18(4):860-875. AbstractWebsite

The article presents a review of the use of cross-section and staining techniques for investigating natural organic materials (mainly proteinaceous and oil-based binders/varnishes) in painted and polychrome artworks, considering the requirements of conservation practice and routine diagnostics. The reviewed literature calls attention to the importance of using cross sections to prepare samples for optical microscopy and to different properties of embedding resins; the most appropriate instrumental conditions for optical microscopy; and the advantages and disadvantages of the most common staining techniques. A few case studies were selected to illustrate the use of autofluorescence (intrinsic fluorescence) and induced fluorescence (using specific staining tests and fluorophore-labeled antibodies) for mapping and identifying organic paint materials in cross sections. New directions of research in cross-section analyses and fluorescence-based techniques for the identification and mapping of artistic materials are presented. The complementary use of different stains on the same cross section, further exploration of intrinsic and induced fluorescence of aged versus fresh materials, and applicability of cross-section observation and staining as complementary methods for assessing the effectiveness of restoration treatments, such as cleaning and consolidation, are discussed in the last section of the article.

Barroso, T, Roque ACA, Aguiar-Ricardo A.  2012.  Bioinspired and Sustainable Chitosan Based Monoliths for Antibody Capture and Release. RSC ADV. 2(30):11285-11294. AbstractWebsite

Chitosan-based monoliths activated by plasma technology induced the coupling of a robust biomimetic ligand, previously reported as an artificial Protein A, with high yields while minimizing the environmental impact of the procedure. Due to the high porosity, good mechanical and tunable physicochemical properties of the affinity chitosan-based monoliths, it is possible to achieve high binding capacities (150 ± 10 mg antibody per gram support), and to recover 90 ± 5% of the bound protein with 98% purity directly from cell-culture extracts. Therefore, the chitosan-based monoliths prepared by clean processes exhibit a remarkable performance for the one-step capture and recovery of pure antibodies or other biological molecules with biopharmaceutical relevance.

Ataíde, F, Azevedo C, Clemente JJ, Cunha AE, Freitas F, Reis MAM, Roque ACA, Oliveira R.  2012.  Analysis of oxygen transport enhancement by functionalized magnetic nanoparticles (FMP) in bioprocesses. New Biotechnology. 29S:S75.Website

Our group was at HUPO 11th Annual World Congress

Íris Batalha was at the HUPO 11th Annual World Congress, that was held in Boston, Massachussetts, USA, from 9 to 13 of September 2012.

Íris presented a Poster, with the title "Novel synthetic scaffolds for the enrichment of phosphorylated proteins and peptides" and won a Travel Grant, provided by the organizers of the Congress.

Borlido, L, Azevedo AM, Sousa AG, Oliveira PH, Roque ACA, Aires-Barros MR.  2012.  Fishing human monoclonal antibodies from a CHO cell supernatant with boronic acid magnetic particles. Journal of Chromatography B. 903:163-170. AbstractWebsite

In this work we have evaluated the potential of boronic acid functionalized magnetic particles for the one-step capture of a human monoclonal antibody (mAb) from a Chinese hamster ovary (CHO) cell culture supernatant. For comparison, Protein A coated magnetic particles were also used. The most important factor influencing the overall process yield and product purity in boronic acid particles was found to be the binding pH. Basic pH values promoted higher purities while resulting in decreased yields due to the competing effects of molecules such as glucose and lactate present in the cell culture supernatant. After optimization, the particles were successfully used in a multi-cycle purification process of the mAb from the CHO feedstock. Boronic acid particles were able to achieve an average overall yield of 86% with 88% removal of CHO host cell proteins (HCP) when the binding was performed at pH 7.4, while at pH 8.5 these values were 58% and 97%, respectively. In both cases, genomic DNA removal was in excess of 97%. Comparatively, Protein A particles recorded an average overall yield of 80% and an HCP removal greater than 99%. The adsorption of the mAb to the boronic acid particles was shown to be mediated by strong affinity interactions. Overall, boronic acid based purification processes can offer a cost-effective alternative to Protein A as the direct capturing step from the mammalian cell culture.

Santana, SDF, Pina AS, Roque ACA.  2012.   Immobilization of enterokinase on magnetic supports for the cleavage of fusion proteins. Journal of Biotechnology. 161:378–382. AbstractWebsite

Magnetic nanobiocatalysts for tag cleavage on fusion proteins have been prepared by immobilizing
enterokinase (EK) onto iron oxide magnetic nanoparticles coated with biopolymers. Two different
chemistries have been explored for the covalent coupling of EK, namely carbodiimide (EDC coupling)
and maleimide activation (Sulfo coupling). Upon immobilization, EK initial activity lowered but EDC coupling lead to higher activity retention. Regarding the stability ofthe nanobiocatalysts,thesewere recycled
up to ten times with the greater activity losses observed in the first two cycles. The immobilized EK also
proved to cleave a control fusion protein and to greatly simplify the separation of the enzyme from the
reaction mixture.

Annual Group Day - Mafra and Ericeira

Our annual group day was on Mafra and Ericeira region! We were not very lucky with the weather, but we still had a great time!