Analytical strategies for detecting nanoparticle–protein interactions

Abstract

A significant increase in biomedical applications of nanomaterials and their potential toxicity demand versatile analytical techniques to determine protein – nanoparticle (NP) interactions. These diverse analytical techniques are reviewed. Spectroscopic methods play a significant role in studying binding affinity, binding ratio, and binding mechanisms. To elucidate NP-proteome interactions, chromatography and electrophoresis techniques are applied to separate NP-bound proteins and Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to identify these proteins. Since NP-protein binding is a dynamic event, Surface Plasmon Resonance (SPR) and Quartz Crystal Microbalance (QCM) are methods of choice to study kinetics of NP-protein binding.

Biomolecular coronas provide the biological identity of nanosized materials

“…The hard corona structure can also be studied by isolating the particle-corona , surface plasmon resonance 32,86,92 , small angle X-ray scattering 67,91,93 , quartz crystal microbalance 94 , isothermal titration calorimetry 86,93,95 , UV-vis and fluorescence spectroscopies 68,96 . Adsorption on the nanoparticle surface may result in protein conformational changes, and these have been studied by circular dichroism 6, 67,68,72,80 , infrared spectroscopy 69,80 , NMR 85,97 , and enzyme activity assays 67,68,98 .…”

Cellulase biocatalysis: key influencing factors and mode of action

“…A third acidic amino acid residue called aspartic acid (D214) has been reported to assist the nucleophilic attack. The mutations of these two active site amino acids have shown the reduction of rate of hydrolysis by 85-370 fold (Barnett et al 2011;Li et al 2010;Yan et al 2011 (Zhang et al 2013). TrCel6A, glycoside hydrolases (GH6 and GH7) TrCel6A is a processive cellulase and emphasized the requirement for accurate GH6 processivity assays (Payne et al 2015) GH6 demonstrated higher hydrolysis activity towards homoglycosidic linkage of cellulose polymers, but relatively poor hydrolysis of the heteroglycosidic linkages Turnover of GH6 of these molecules was remained slow when compared to GH7 activity Fig.…”

Möglichkeiten und Limitierungen verschiedener Trenntechniken zur Analyse der Proteinkorona

“…Die treibende Kraft fürd ie Trennung ist die Bewegung durch ein elektrisches Feld. [15] Anhand dieser Einblicke in die Proteinkorona wird deutlich, dass CE eine vielversprechende Te chnik auf dem Gebiet der Abtrennung der Proteinkorona ist und dass die isolierten Protein-Nanoträger-Komplexe zur weiteren Analyse verwendet werden kçnnen. Durch die spezifische Wanderungszeit konnten die Autoren Konjugate verschiedener AuNCs mit einzelnen Proteinen identifizieren.…”