Ana Rita Santos

Class 2017phdARS

 

PhD thesis

Biomanufacturing of DNA-origami nanostructures for bio-imaging

 

Supervisor

• Miguel F. Prazeres, Professor @ IST/ULisboa, PT

• Pedro Paulo, Research Scientist @ IST/ULisboa, PT

Collaborators

• Hendrik Ditz, Professor @ TUMünchen, Munique, D

 

Proposal summary

DNA nanostructures assembled from large (~500-50000 bp) single stranded DNA scaffolds and short staple strands via DNA origami techniques are finding application as components of nanomachines, nanorobots and biosensors. Currently, scaffolds are obtained by asymmetric PCR or from the DNA of phages like M13, and purified using a range of lab-based, cumbersome techniques that are neither scalable nor compatible with drug delivery and biosensing applications. The goals of this project are (i) to develop scalable processes for the biomanufacturing of scaffolds and DNA-origami nanostructures, and (ii) to study the possibility of using fluorescently labelled DNA nanostructures in bio-imaging applications. Scaffolds will be purified from PCR mixtures or from the M13 DNA using scalable operations like precipitation and chromatography. Nanostructures will be assembled from these scaffolds and purified further. The obtained nanostructures are labeled with fluorophores and studied by FRET and confocal fluorescence lifetime microscopy techniques.

 

PUBLICATIONS

Papers

Silva-Santos AR, Alves CPA, Monteiro M, Azevedo AM, Prazeres DMF, Multimodal chromatography of supercoiled minicircles: A closer look into DNA-ligand interactions, Separation and Purification Technology, 212, 161-170, 2019.

 

Oral Communications

Capela EV, Silva-Santos AR, Campos-Pinto I, Rodríguez MA, Gavara PR, Fernandez-Lahore M, Aires-Barros MR, Azevedo AM, “LYTAG-driven purification strategies as a key to integrate and intensify the downstream processing of monoclonal antibodies”, MicroBiotec2017, December 7-8, 2017, Porto, Portugal

Silva-Santos AR, Prazeres DMF, “Chromatographic purification of single stranded DNA scaffolds for biomanufacturing DNA-origami nanostructures”, MicroBiotec2019, December 5-7, 2019, Coimbra, Portugal

 

Poster Communications

Silva-Santos AR, Alves CPA, Šimčiková M, Monteiro G, Azevedo AM, Prazeres DMF, “Multimodal chromatography: a closer look to supercoiled minicircle DNA interactions”, MicroBiotec2017 December 7-8, 2017, Porto, Portugal

Silva-Santos AR, Prazeres DMF, “Biomanufacturing of DNA-origami nanostructures for bio-imaging”, PhD open days 2018Instituto Superior Técnico, Lisboa, Portugal

A.Rita Silva-Santos, Cláudia P.A. Alves, Michaela Simčíková, Gabriel Monteiro, Ana M Azevedo, Duarte Miguel F. Prazeres, “Multimodal chromatography of supercoiled minicircles: a closer look into DNA-ligand interactions, 12th ESBES – European Symposium on Biochemical Engineering Sciences, September 2018, Lisboa, Portugal

 

DOCTORAL PROGRAM (36 ECTS)

• General Doctoral Training (6 ECTS)

• Advanced Experimental Techniques and methodologies (6 ECTS)

• Bioentrepreneurship (6 ECTS)

  • Outreach and Teaching Skills (6 ECTS)

  • Nanobiotechnology (6 ECTS)

  • Advanced Topics in Bioengineering and Biosciences (6 ECTS)


Advanced courses:

• ABC2 – Anything but conventional chromatography workshop, Lisbon, Portugal, November 22-24, 2017

• BIOPRO World Talent Campus, organized by DTU and sponsored by NovoNordisk Foundation, Copenhagen, Denmark, August, 2018

• Cell Culture-based Viral Vaccines course – 3rd edition, organized by the European Society for Animal Cell Technology in Llafranc, Girona, Spain, September, 2018

• Drug Development course – 3rd edition, organized by the European Society for Animal Cell Technology in Llafranc, Girona, Spain, September, 2018