Microfluidics-based LC-MS MRM approach for the relative quantification of Burkholderia cenocepacia secreted virulence factors.

Burkholderia cenocepacia is an opportunistic pathogen that is commonly isolated from patients with cystic fibrosis (CF). Several virulence factors have been identified, including extracellular enzymes that are secreted by type II and type VI secretion systems. The activity of these secretion systems is modulated by quorum sensing. Apart from the classical acylhomoserine lactone quorum sensing, B. cenocepacia also uses the diffusible signal factor system (DSF) i.e. 2-undecenoic acid derivatives that are recognized by specific receptors resulting in changes in biofilm formation, motility and virulence. However, quantitative information on alterations in the actual production and release of virulence factors upon exposure to DSF is lacking. We here describe an approach implementing microfluidics based chromatography combined with single reaction monitoring to quantify protein virulence factors in the secretome of B. cenocepacia.