Nanomechanics on FGF-2 and Heparin Reveal Slip Bond Characteristics with pH Dependency.

Fibroblast growth factor 2 (FGF-2), an important paracrine growth factor, binds electrostatically with low micromolar affinity to heparan sulfates present on extracellular matrix proteins. A single molecular analysis served as a basis to decipher the nanomechanical mechanism of the interaction between FGF-2 and the heparan sulfate surrogate, heparin, with a modular atomic force microscope (AFM) design combining magnetic actuators with force measurements at the low force regime (1 × 10 to 1 × 10 pN/s). Unbinding events between FGF-2-heparin complexes were specific and short-lived.

Dually actuated atomic force microscope with miniaturized magnetic bead-actuators for single-molecule force measurements.

We report a novel atomic force microscopy (AFM) technique with dual actuation capabilities using both piezo and magnetic bead actuation for advanced single-molecule force spectroscopy experiments. The experiments are performed by manipulating magnetic microbeads using an electromagnet against a stationary cantilever. Magnetic actuation has been demonstrated before to actuate cantilevers, but here we keep the cantilever stationary and accomplish actuation via free-manipulated microstructures.

Probing unnatural amino acid integration into enhanced green fluorescent protein by genetic code expansion with a high-throughput screening platform.

Background: Genetic code expansion has developed into an elegant tool to incorporate unnatural amino acids (uAA) at predefined sites in the protein backbone in response to an amber codon. However, recombinant production and yield of uAA comprising proteins are challenged due to the additional translation machinery required for uAA incorporation.

Results: We developed a microtiter plate-based high-throughput monitoring system (HTMS) to study and optimize uAA integration in the model protein enhanced green fluorescence protein (eGFP).

Drug delivery of Insulin-like growth factor I.

This review starts off outlining the control of Insulin-like growth factor I (IGF-I) kinetics in Nature and by virtue of a complex system of 6 binding proteins controlling half-life and tissue distribution of this strong anabolic peptide. In addition, alternative splicing is known to result in IGF-I variants with modulated properties in vivo and this insight is currently translated into advanced IGF-I variants for therapeutic use. Insights into these natural processes resulted in biomimetic strategies with the ultimate goal to control pharmacokinetics and have recently propelled new developments leading to optimized pharmaceutical performance of this protein in vivo.