Multiplexed Probing for Functional Capacity Measurements in Complex and Limited-Size Samples
EMSL Project ID
50578
Abstract
The TIP LDRD program at PNNL has supported the development, application and patenting of novel strategies to functionalize activity-based chemical probes to glass surfaces for making measurements of functional capacity in complex biological samples. The final step in this effort is pairing the activity-based probes with fluorescent glass microspheres, which will enable the use of probes for multiple different enzyme targets in a single limited-size sample with tandem direct quantification of target enzymes using Fluorescence-Activated Cell Sorting (FACS) and proteomics. Making functional measurements in biological samples suffers from setbacks, particularly limited sample size and physiological settings incompatible with traditional chemical probing strategies. To remedy this, we propose an approach to provide layers of functional resolution: (1) demonstrate probe functionalization and pairing to fluorescent glass microspheres, and then multiplex the probe-functionalized microspheres in limited size samples to label target enzymes. (2) The amount of enzyme targets in a given sample can first be quantified by FACS, and subsequently the specific targets can be identified and quantified by mass spectrometry-based proteomics. In the near future kits can be prepared that contain one or more probe type given the end users’ needs. Once successfully commercialized, this multiplexed assay can be used with little to no technical expertise, yielding broad potential licensing opportunities.
Project Details
Start Date
2018-10-29
End Date
2019-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members