Impact of Nanoparticle Aggregation on Protein Recovery through a Pentadentate Chelate Ligand on Magnetic Carriers

The growing need for more efficient separation techniques still dominates downstream processing of biomolecules, thus encouraging the continuous development of advanced nanomaterials. In this paper we present an improved process for recovering recombinant histidine tagged green fluorescent protein from an <i>E. coli</i> cell lysate. Superparamagnetic core–shell nanocarriers are functionalized with a pentadentate chelate affinity ligand and then loaded with metal ions (Cu²⁺, Ni²⁺, or Zn²⁺). The separation process yields high binding capacity (250 mg/g), good selectivity, purity >98%, good recyclability with 90% capacity after 9 cycles, and long-term stability. We determined the main physical properties of the magnetite-based nanoparticles such as saturation magnetization (59 A m²/kg), primary particle diameter (22 ± 4 nm), and specific surface area (89 m²/g). Our results show that this material is a promising tool for bioseparation applications. One special focus of the work includes analyzing the changes in the hydrodynamic size distribution using dynamic light scattering and transmission electron microscopy. We relate these effects to different interaction levels in the system and discuss how the stronger aggregation of the magnetite core is the main limiting factor for the separation yield, leading to a considerable decrease in the number of metal ions available for biomolecular capture. Otherwise weaker interactions lead instead to agglomeration effects that have no impact on the binding capacity of the system. The simple relation between the size of the aggregated units and the size of the primary particles corresponds approximately to the relation between the number of existing binding sites and the actual protein binding in the separation process. Compared with that, the effect of steric hindrance among proteins is of less significance

All-in-One Nanowire-Decorated Multifunctional Membrane for Rapid Cell Lysis and Direct DNA Isolation

[Image: see text] This paper describes a handheld device that uses an all-in-one membrane for continuous mechanical cell lysis and rapid DNA isolation without the assistance of power sources, lysis reagents, and routine centrifugation. This nanowire-decorated multifunctional membrane was fabricated to isolate DNA by selective adsorption to silica surface immediately after disruption of nucleus membranes by ultrasharp tips of nanowires for a rapid cell lysis, and it can be directly assembled with commercial syringe filter holders. The membrane was fabricated by photoelectrochemical etching to create microchannel arrays followed by hydrothermal synthesis of nanowires and deposition of silica. The proposed membrane successfully purifies high-quality DNA within 5 min, whereas a commercial purification kit needs more than an hour