Proof of concept of a "greener" protein purification/enrichment method based on carboxylate-terminated carbosilane dendrimer-protein interactions.
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.
Departamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871, Alcalá de Henares, Madrid, Spain.
Estefanía
E
González-García
Faculty of Science, J. E. Purkinje University, Ceske mladeze 8, 40096, Usti nad Labem, Czech Republic.
Marek
M
Maly
Departamento de Química Orgánica y Química Inorgánica, Universidad de Alcalá, Ctra. Madrid-Barcelona, Km 33.600, 28871, CIBER-BBN, Alcalá de Henares, Madrid, Spain.
Francisco Javier
FJ
de la Mata
Departamento de Química Orgánica y Química Inorgánica, Universidad de Alcalá, Ctra. Madrid-Barcelona, Km 33.600, 28871, CIBER-BBN, Alcalá de Henares, Madrid, Spain.
Rafael
R
Gómez
Departamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871, Alcalá de Henares, Madrid, Spain.
María Luisa
ML
Marina
Departamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871, Alcalá de Henares, Madrid, Spain. concepcion.garcia@uah.es.
María Concepción
MC
García
D002264:Carboxylic Acids / Q000737:chemistry*; D011232:Chemical Precipitation; D050091:Dendrimers / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D056004:Molecular Dynamics Simulation; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification*; D009211:Myoglobin / Q000737:chemistry / Q000302:isolation & purification*; D010940:Plant Proteins / Q000302:isolation & purification; D011485:Protein Binding; D017433:Protein Structure, Secondary; D000068242:Prunus domestica / Q000737:chemistry; D012639:Seeds / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification*; D012821:Silanes / Q000737:chemistry*; D012997:Solvents
D002264:Carboxylic Acids; D050091:Dendrimers; D009211:Myoglobin; D010940:Plant Proteins; D012821:Silanes; D012997:Solvents; C504072:carbosilane; D012710:Serum Albumin, Bovine; D009113:Muramidase
Carboxylate-terminated carbosilane dendrimers; Computer modeling; Fluorescence quenching; Molecular dynamics; Protein sample preparation; Protein-dendrimer interaction