Biacore will be presenting a scientific poster demonstrating the application of automated SPR-MS at the International Mass Spectrometry Conference in Edinburgh
Biacore International, in collaboration with Bruker Daltonics, will be presenting a scientific poster demonstrating the application of automated SPR-MS as a tool to identify and analyse function of proteins.
The poster demonstrates the use of Biacore3000 surface plasmon resonance (SPR) based system in combination with mass spectrometry (MS) to solve a typical proteomics problem to identify interaction partners of a protein obtained via ligand fishing.
Biacore3000 uniquely provides a fully automated workflow, which includes binding of the analyte, recovery, optional on-target digestion, and preparation of the sample for a number of MS techniques including, Maldi-ToF, Maldi ToF/ToF MS/MS and LC-ESI MS/MS.
Using calmodulin and the calmodulin-binding domain (CBD) of myosin light chain kinase as a model system, the poster explains how analyte recovery, with direct sample deposition onto the Maldi plate, and SPR-MS experiments are carried out with the help of dedicated software wizards.
Calmodulin, a ubiquitous Ca-binding protein which serves as a major physiological effector for a wide range of cellular responses, was captured from bovine brain extract on a CBD-modified chip.
From this crude brain extract mixture, the fished protein was recovered in a highly purified state and identified directly using Maldi-ToF MS.
Further experiments were also carried out using LC-ESI MS/MS to sequence the recovered and digested calmodulin material from the original ligand fishing experiment.
"Integration of Biacore3000 with mass spectrometry certainly represents an extremely powerful tool for function-based molecular characterisation and identification," said Clive Seymour, head of Biacore's life science research business unit.
"In addition to this poster, there are now a number of papers published that demonstrate the effectiveness of this powerful technology combination approach".
The enhanced Biacore3000 functionality has also been field tested as part of collaborations in this field with laboratories in a number of key organisations worldwide.
