Disruptive effects of protein on performance of liquid membrane-based ion-selective microelectrodes.

The effects of addition of protein [bovine serum albumin (BSA)] to the calibration solutions of various liquid membrane-based ion-selective microelectrodes have been assessed. Protein at typically cytosolic levels of 100 mg/ml had no effect on the response of Na(+)-, K(+)-, Mg(2+)-, Ca(2+)-, H(+)-, NO3(-)-, and Cl(-)- selective microelectrodes at ion concentrations well above the respective detection limits. By contrast, this level of protein resulted in a pronounced increase in the detection limits of Na(+)-, K(+)-, Mg(2+)-, Ca2+, and Cl(-)-selective microelectrodes; H+ and NO3- microelectrodes were largely unaffected by the protein. The effect of BSA on detection limit is independent of pH and ionic strength, nor is it due to hysteresis, to binding of ions to the protein, or to the generation of junction potentials. More extensive studies on Na(+)-selective microelectrodes revealed that other proteins and large molecules (e.g., polyethylene glycol) also increase detection limits, with the extent of this effect apparently correlated positively with the molecular weight. Because biological macromolecules at physiological concentrations can influence the properties of alkali- and alkali earth cation-selective microelectrodes, care must be taken if the use of these electrodes entails cytosolic measurements close to their detection limits.