Development of native two-dimensional electrophoresis methods for the separation and detection of platinum carrying serum proteins: initial steps

Abstract

The initial development steps of a native and powerful two-dimensional electrophoretic (2-D) method for the separation of platinum-proteins is described. Mild conditions were selected, particularly for the second dimension, e.g., avoiding buffer systems with platinophile N- or S-donor groups. Therefore, the separation reagents were checked if and at which concentration they can be used for this purpose. In the first dimension isoelectric focusing (IEF) was performed using immobilised pH gradients (IPGs). Native polyacrylamide gel electrophoresis (PAGE) was done in the second dimension. Detection of proteins was achieved via silverstaining. For the determination of platinum in the ultra-trace range, double focusing inductively coupled plasma mass spectrometry (HR-ICP-MS) was used. Autoradiography (¹⁹¹Pt tracer) will be done additionally in the future as a fast, powerful and elegant way of detecting the platinum carrying proteins after the second dimension.