Protein charge organization is dependent on the low-permittivity region in the hydrophobic core of the molecule. We suggest a novel approach to estimate the dielectric constant of this region by comparing measured and simulated first- and second-order charge moments. Here, the dipole moment is measured as a function of pH using dielectric spectroscopy. The results are compared to dipole moments based on Poisson-Boltzmann estimates of pK(a) shifts calculated from structures in the Protein Data Bank. Structures are additionally refined using CHARMM molecular dynamics simulations. The best estimate for the internal permittivity is found by minimizing the root-mean-square residual between measured and predicted charge moments. Using the protein β-lactoglobulin, a core dielectric constant in the range of 6-7 is estimated.

A liquid impedance cell for broadband impedance measurements up to 110 MHz is presented. The design incorporates temperature control and minimizes parasitic capacitance and inductance. The cell is simple to fabricate and uses chemically resistant materials, stainless steel, and Teflon. This dielectric cell can be used in a variety of liquid measurements, particularly those related to impedance measurements of biological objects in solution. Temperature control is illustrated in measurements of the permittivity of deionized water from 5 to 55  °C. Numerical fitting procedures employed on the relaxation curves indicate good agreement with previous studies on beta-lactoglobulin and hen lysozyme. Titration capability is demonstrated through dielectric titration of hen lysozyme and beta-lactoglobulin.

Electrode polarization is a significant obstacle in the impedance measurements of ionic liquids. An atomically smooth electrode surface could potentially reduce unwanted impedance contributions from electrode polarization. Liquid metal electrodes were formed by adhering Galinstan to acrylic plates in a parallel-plate capacitor arrangement. Electrode polarization was compared to a similar cell with stainless steel electrodes. The impedance of salt and protein solutions (β-lactoglobulin) was measured from 40 Hz to 110 MHz. Because of oxide layer formation, the performance of the Galinstan electrode is significantly different than the theoretical ideal.

Dielectric spectroscopy measurements of liquids are often limited by electrode polarization. The influence of surface polishing and deposition of the conducting polymer polypyrrole/polystyrenesulfonate (PPy/PSS) on the polarization impedance is investigated. A quantitative description of the electrode polarization contribution to the real-valued permittivity spectrum is derived. This description explains the origin of the ω(-const). (const.>1) dependency commonly observed in permittivity measurements. Electrode surface roughness is correlated with both the magnitude and phase of the constant phase element. Generally, rougher electrodes have better performance, and an order of magnitude bandwidth improvement is achieved using PPy/PSS electrodes.

This paper presents a quantitative approach for measuring pH-controlled protein aggregation using dielectric spectroscopy. The technique is demonstrated through two aggregation experiments, the first between β-lactoglobulin (β-Lg) and hen lysozyme (HENL) and the second between bovine serum albumin (BSA) and HENL. In both experiments, the formation of aggregates is strongly dependent on the solution pH and is clearly indicated by a decrease in the measured permittivity when the second protein is added. A quantifiable lower-bound on the ratio of proteins involved in the aggregation process is obtained from the permittivity spectra. Lower-bound aggregation ratios of 83 % for β-Lg/HENL at pH 6.0 and 48 % for BSA/HENL at pH 9.2 were consistent with turbidity measurements made on the same solutions.