Toward High-throughput, Quantitative Platforms to Identify the Targets of Small Molecules

Target identification is a major challenge in probe and drug discovery. Current binding assays are unable to detect interactions between unoptimized probes and difficult targets, such as transcription factors. Here, we developed generalizable, high-throughput platforms that can rapidly identify the mechanism(s) of action of small molecules emerging from high-throughput screening (HTS) campaigns. Specifically, this project established a solid phase method to rapidly modify small molecules with moieties of interest using isocyanate-based chemistries. This chemical method can be used to quickly generate photoaffinity labeling analogs of small molecules that can be used in a covalent ELISA and mass spectrometry workflow to determine whether small molecules bind to a target of interest and identify off-target binders. Additionally, we created a synergistic critical path for assessing the mechanism of action and on-target activity of small molecules through the generation of an on-target transcriptional profile and application of the L1000 gene-expression platform. Together, these workflows and chemical tools will enable high-throughput studies of small molecule-protein interactions with a wide range of affinities and abundances and facilitate prioritization of small molecules that bind and modulate the function of difficult targets.

Name thesis:degree_name

Doctoral

Department dc:contributor.department

Massachusetts Institute of Technology. Department of Biological Engineering

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2023

Author dc:creator

• Henry, Catherine Campbell

Advisor dc:contributor.advisor

• Koehler, Angela N.

Statement dc:rights

• In Copyright - Educational Use Permitted
• Copyright MIT

Licence dc:rights.uri

• http://rightsstatements.org/page/InC-EDU/1.0/

Handle dc:identifier.uri

https://hdl.handle.net/1721.1/150178

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/150178