EpiProfileLite

EpiProfileLite profiles approximately 200 histone post-translational modifications (PTMs) from tissue or cell line samples using direct infusion mass spectrometry (DI-MS) to enable rapid quantitative analysis of histone modification states.

Key Features:

• Direct Infusion Mass Spectrometry (DI-MS): Uses DI-MS with direct injection enabling ~1 minute MS acquisition per sample and obviating nano-liquid chromatography (nanoLC).
• Histone PTM Coverage: Quantifies approximately 200 histone PTMs from tissue or cell line samples.
• Sample Preparation and Parallelization: Employs a protocol with ~4-hour histone extraction and ~3-hour derivatization and digestion and supports parallel processing of up to 384 samples using multichannel pipettes and 96-well plates.
• Quality Assurance: Incorporates a synthetic "histone-like" peptide spike-in to benchmark derivatization and digestion and compares DI-MS results to nanoLC-MS/MS using a library of synthetically modified histone peptides to assess ionization bias.
• Throughput: Capable of processing over 1,000 samples per day on a single mass spectrometer.

Scientific Applications:

• Chromatin dynamics and epigenetics: Provides comprehensive histone PTM profiles to study chromatin dynamics and epigenetic regulation.
• Chromatin accessibility and gene expression: Aids investigation of how histone PTMs influence chromatin accessibility and gene expression regulation.
• Enzyme recruitment and remodeling: Supports studies of recruitment of enzymes responsible for DNA readout and chromatin remodeling.
• Transcription and DNA repair: Enables analysis of histone modification changes associated with transcriptional activation, repression, and DNA repair.

Methodology:

The workflow integrates histone extraction, derivatization, digestion, and direct infusion mass spectrometry.