0000000000424153
AUTHOR
Maximilian Brückner
Bio-orthogonal triazolinedione (TAD) crosslinked protein nanocapsules affect protein adsorption and cell interaction
Albumin-based protein nanocarriers have been widely exploited as drug delivery systems, since they show excellent degradability, low toxicity, but at the same time provide high loading capacity and relevant uptake into cells. For the formation of protein nanocapsules, bio-orthogonal reactions are important so that the material to be encapsulated is not affected by the shell formation. We show that protein nanocapsules with narrow size distributions and low protein adsorption upon contact with blood serum can be synthesized by inverse miniemulsion and interfacial crosslinking of the protein using triazolinediones (TADs) as powerful dienophiles and enophiles, which smoothly perform electrophi…
Preparation of the Protein Corona: How Washing Shapes the Proteome and Influences Cellular Uptake
A protein coat, termed the protein corona, assembles around the nanocarriers´ surface once it gets in contact with a biological environment. We show that the media used for washing of the protein corona can be crucial. This is true for downstream analysis as well as for precoating for in vitro or in vivo use. This has been widely overlooked so far. We focus on the choice of eight different washing media and how they influence the composition of the hard protein corona of several nanocarriers incubated with human blood plasma and serum. SDS-PAGE and LC-MS analysis showed major differences in protein corona profiles when using diverse washing media. While plasma and serum proteins already hav…
Monitoring of Cell Layer Integrity with a Current-Driven Organic Electrochemical Transistor
Abstract The integrity of CaCo-2 cell barriers is investigated by organic electrochemical transistors (OECTs) in a current-driven configuration. Ion transport through cellular barriers via the paracellular pathway is modulated by tight junctions between adjacent cells. Rupturing its integrity by H2O2 is monitored by the change of the output voltage in the transfer characteristics. It is demonstrated that by operating the OECT in a current-driven configuration, the sensitive and temporal resolution for monitoring the cell barrier integrity is strongly enhanced as compared to the OECT transient response measurement. As a result, current-driven OECTs are useful tools to assess dynamic and crit…
The conjugation strategy affects antibody orientation and targeting properties of nanocarriers.
Antibody-modified drug delivery systems in the nano-range have the ability to overcome current challenges for treating diseases due to their high specificity towards the targeted body region. However, no antibody-bound nanocarrier has been clinically approved to date. This missing clinical approval may be a result of the conjugation strategy that influences the spatial orientation of the attached antibody on the nanocarriers' surface. What is not missing, however, is a diverse selection of antibody to nanocarrier conjugation strategies that determine the success of an antibody functionalized drug delivery system. In this paper, two antibody conjugation strategies were compared by conjugatin…
Current-Driven Organic Electrochemical Transistors for Monitoring Cell Layer Integrity with Enhanced Sensitivity
In this progress report an overview is given on the use of the organic electrochemical transistor (OECT) as a biosensor for impedance sensing of cell layers. The transient OECT current can be used to detect changes in the impedance of the cell layer, as shown by Jimison et al. To circumvent the application of a high gate bias and preventing electrolysis of the electrolyte, in case of small impedance variations, an alternative measuring technique based on an OECT in a current-driven configuration is developed. The ion-sensitivity is larger than 1200 mV V-1 dec-1 at low operating voltage. It can be even further enhanced using an OECT based complementary amplifier, which consists of a p-type a…