Probing Biomolecular Recognition at the Single Molecule Level

Sammanfattning: Specific recognition between biomolecular partners contributes to a multitude of biochemical signaling within and between cells. The aim of this work has been to investigate the possibility to probe biomolecular recognition reactions at the smallest possible scale, the single molecule level. The ability to do so provides unique possibilities to understand and acquire information of biological systems utilizing lower analyte concentrations and/or obtain information hidden in ensemble measurements. By exploring several of the unique properties of liposomes, we have in this work developed assay formats capable of analyzing individual molecular interactions. Taking advantage of the fact that liposomes are compatible with the important class of cell membrane-residing proteins, we investigated binding kinetics of ligand interactions with membrane receptors. Monitoring individual binding events also provides possibilities for parallel detection of multiple interaction partners. In exploring this aspect, we have taken the first steps in the development of a new means to perform multiplexed biorecognition analysis using imaging mass spectrometry. Finally, future and ongoing extensions of the liposome-based assay are presented, illustrating two clinically relevant applications, screening for virus detection and mass spectrometry-based immunohistochemistry.