Detection of Multiple Proteins on One Spot by Laser Ablation Inductively Coupled Plasma Mass Spectrometry and Application to Immuno- Microarray with Element-Tagged Antibodies

Abstract

Inductively coupled plasma mass spectrometry (ICPMS) has been successfully used for the detection of element-tagged biomolecules with the advantage of multielement capability. However, this technique cannot be used for microarray detection due to the necessity to dissolve the elemental tags before introducing them to the plasma source. Here, we report the detection of multiple proteins on each spot of the immuno-microarray by laser ablation ICPMS. alpha-Fetoprotein IgG (AFP), carcinoembryonic antigen (CEA), and human IgG, as model proteins, have been detected on the basis of sandwich-type immunoreactions on a microarray with Sm3+-labeled AFP antibody, Eu3+-labeled CEA antibody, and Au-labeled goat-anti-human IgG (GAH) as labeled antibodies. The detection limits were 0.20, 0.14, and 0.012 ng mL-1 (3sigma) with the RSD of 5.7%, 2.6%, and 2.3% at the concentration of 1.0 ng mL-1 for AFP, CEA, and human IgG, respectively. The present detection method permits detecting multiple analytes from each spot of microarray with a spatial resolution at micrometer range, which can alleviate the stress to fabricate high-density arrays. Furthermore, the substrate materials and immobilized proteins do not interfere with the detection. The present technique provides a new strategy for readout of microarray.