Utilization of albumin-based sensor chips for the detection of metal content and characterization of metal-protein interaction by surface plasmon resonance

We report here the use of albumin-based biosensor chips for the determination of metal content and characterization of metal-protein interaction by surface plasmon resonance. Bovine serum albumin was immobilized onto a carboxymethylated dextran matrix and used for metal detection. The temperature for the analysis was defined and the highest interaction was observed at 25 °C. The albumin sensor chip binds cadmium, zinc or nickel in a concentration-dependent manner, but not magnesium, manganese and calcium. The optimal buffer condition used for the analysis contains 0.01 M HEPES, pH 7.4, 1 mM NaCl and 0.005% Tween-20. Using this condition, a linear calibration curve within the range of 10−8 to 10−4 M can be established for the metals. However, a dramatic increase in binding capacity was observed when metal concentration was higher than 10−4 M and reached a plateau at 10−2 M. The detection limit for Cd can reach as low as 1 ppb. When measuring a solution containing two species of metal ions with the albumin chip, an additive effect was observed for Ni and Zn. However, 20-30% reduction in resonance response was found upon mixing Cd with Zn or Ni. These observations are consistent with the binding characteristics of albumin. The feasibility of measuring serum metal content by the albumin chip was examined. A linear calibration curve can be established if the samples are boiled and passed through a gel filtration column. The binding affinity of metal with albumin can also be achieved by using the sensor chip. The binding affinity follows the order of Ni > Zn > Cd. These results indicate that the albumin-based sensor chip is useful not only in the quantification of metal content, but also in the characterization of the biochemical properties of albumin.