Analytical and Bioanalytical Chemistry

, Volume 408, Issue 27, pp 7679–7687 | Cite as

Proof of concept of a “greener” protein purification/enrichment method based on carboxylate-terminated carbosilane dendrimer-protein interactions

  • Estefanía González-García
  • Marek Maly
  • Francisco Javier de la Mata
  • Rafael Gómez
  • María Luisa Marina
  • María Concepción García
Research Paper


Protein sample preparation is a critical and an unsustainable step since it involves the use of tedious methods that usually require high amount of solvents. The development of new materials offers additional opportunities in protein sample preparation. This work explores, for the first time, the potential application of carboxylate-terminated carbosilane dendrimers to the purification/enrichment of proteins. Studies on dendrimer binding to proteins, based on protein fluorescence intensity and emission wavelengths measurements, demonstrated the interaction between carboxylate-terminated carbosilane dendrimers and proteins at all tested pH levels. Interactions were greatly affected by the protein itself, pH, and dendrimer concentration and generation. Especially interesting was the interaction at acidic pH since it resulted in a significant protein precipitation. Dendrimer-protein interactions were modeled observing stable complexes for all proteins. Carboxylate-terminated carbosilane dendrimers at acidic pH were successfully used in the purification/enrichment of proteins extracted from a complex sample.

Graphical Abstract

Images showing the growing turbidity of solutions containing a mixture of proteins (lysozyme, myoglobin, and BSA) at different protein:dendrimer ratios (1:0, 1:1, 1:8, and 1:20) at acidic pH and SDS-PAGE profiles of the corresponsing supernatants. Comparison of SDS-PAGE profiles for the pellets obtained during the purification of proteins present in a complex sample using a conventional “no-clean” method based on acetone precipitation and the proposed “greener” method using carboxylate-terminated carbosilane dendrimer at a 1:20 protein:dendrimer ratio


Carboxylate-terminated carbosilane dendrimers Protein sample preparation Protein-dendrimer interaction Fluorescence quenching Computer modeling Molecular dynamics 



This work was supported by the Ministry of Economy and Competitiveness (ref. AGL2012-36362 and CTQ-2014-54004-P), the Comunidad de Madrid and European funding from FEDER program (ref. S2013/ABI-3028, AVANSECAL), and Consortium NANODENDMED ref. S2011/BMD-2351 (CAM). The financial support of the Czech Science Foundation (project no. GA15-05903S) is also acknowledged. E.G.-G. thanks the University of Alcalá for her pre-doctoral contract. CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund.

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest to declare.

Supplementary material

216_2016_9864_MOESM1_ESM.pdf (1.5 mb)
ESM 1 (PDF 1498 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Estefanía González-García
    • 1
  • Marek Maly
    • 2
  • Francisco Javier de la Mata
    • 3
  • Rafael Gómez
    • 3
  • María Luisa Marina
    • 1
  • María Concepción García
    • 1
  1. 1.Departamento de Química Analítica, Química Física e Ingeniería QuímicaUniversidad de AlcaláAlcalá de HenaresSpain
  2. 2.Faculty of ScienceJ. E. Purkinje UniversityUsti nad LabemCzech Republic
  3. 3.Departamento de Química Orgánica y Química InorgánicaUniversidad de Alcalá, Ctra. Madrid-BarcelonaAlcalá de HenaresSpain

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