Label-free parallel screening of combinatorial triazine libraries using reflectometric interference spectroscopy.
The parallel reflectometric interference spectroscopy is presented as a label-free optical detection method. A new setup was adapted to accommodate sample carriers in a 96-well microplate. It allows for the first time simultaneous plate imaging by a CCD camera for the parallel detection of specific biomolecular interaction in the microplate wells at heterogeneous phase using direct optical monitoring. The detection of binding events with time resolution enables a highly parallel functional biomolecular interaction analysis (BIA). The combination of this new screening setup with combinatorial solid-phase synthesis is performed in the wells of glass-bottom microplates to accomplish the synthesis and the screening platform within one device. As a model system for a solid-phase substance library, synthesis of a triazine library and the subsequent BIA with four different antibodies were carried out. The presented setup enables a time resolution of 18 s with a total screening time of less than 35 min including baseline adjustment, BIA, and regeneration of the screening device for 96 samples in parallel. The binding studies reveal a fast classification of the different monoclonal and polyclonal antibodies and enable the detection of triazines with high binding affinity. The presented prototype is the first parallelized optical label-free detection system for biomolecular interaction analysis that is suitable for a high-throughput screening based on the 96-well microplate format.
Birkert
Oliver
O
Institute of Physical Chemistry, University of Tübingen, Germany. oliver.birkert@ipc.uni-tuebingen.de
Tünnemann
Rolf
R
Jung
Günther
G
Gauglitz
Günter
G
D000906:Antibodies / Q000737:chemistry; D020650:Combinatorial Chemistry Techniques; D007202:Indicators and Reagents; D008962:Models, Theoretical; D013053:Spectrophotometry; D014227:Triazines / Q000737:chemistry*
D000906:Antibodies; D007202:Indicators and Reagents; D014227:Triazines