Multiplex profiling of tumor‐associated proteolytic activity in serum of colorectal cancer patients

Abstract

Functional protease profiling with RPs might improve the laboratory-based diagnosis, monitoring and prognosis of malignant disease, and has to be evaluated thoroughly in future studies.

Renal clearable catalytic gold nanoclusters for in vivo disease monitoring

“…Monitoring protease activity as a biomarker of disease may be leveraged to overcome the lack of sensitivity and specificity of abundance-based blood biomarkers. 7 Common tools to measure protease activity often rely on cumbersome and infrastructure heavy analyses, such as fluorescence, [8][9][10] mass spectrometry, 11 or MRI. 12 Previously, we developed exogenously administered multiplexed protease-responsive nanoparticles that release small reporter probes into the urine in response to proteolytic cleavage in disease environments.…”

Peptide probes for proteases – innovations and applications for monitoring proteolytic activity

“…Many routes to discover optimal substrates/spacers for FRET protease assays exist. These include a variety of combinatorial approaches (both solid 21,22 and solution based, 23 MS substrate profiling methods, 24,25 phage libraries 26 or substrate modification of known substrates, including direct screening on complex tumour tissues. 27 Many FRET peptide probes have been developed [28][29][30] and there are reviews covering this area so here we highlight some key and novel examples 14,19 with FRET based probes developed for legumain, 31 thrombin, 30 SARS-CoV protease, 32 Granzyme B 33 and Cathepsins, 34 among others.…”

“…The tag-containing peptides co-eluted in chromatographic separations, while the combination of reporter peptides could be analysed by MS/MS. Findeisen developed a series of synthetic MS tagged reporter peptides 25,241,242 to help address a key challenge of protease profiling in complex biological samples, namely the risk of cleavage by non-specific peptidases, that are generally very abundant in complex samples. This was achieved by incorporating a flanking aminohexanoic acid groups in the peptide substrate that successfully reduced degradation by native exopeptidases.…”

Clinical applications of MS‐based protein quantification

“…Finally, not only endogenous plasma proteins can be targeted for diagnostic use. Alternatively, the in vitro processing of synthetic peptides by disease‐related proteases can be monitored using MS‐based quantification .…”