Week 12 — Imaging and Measurement

Date: Tue Apr 22. Slides: Lecture Slides Recording: Class Recording

Info Homework this week is based on data generated at Waters Immerse Cambridge. You’ll characterize eGFP structure (intact mass; peptide map) using LC‑MS. Data will be provided online for remote students. Final Project Homework Warning Mandatory for MIT/Harvard; optional for Committed Listeners (edited Apr 23). Identify at least one aspect of your project you will measure (e.g., protein mass/sequence, biomarker presence/quantity). Describe all elements you will measure and how you will perform these measurements. Specify the technologies you will use (e.g., gel electrophoresis, DNA sequencing, mass spectrometry). Waters Homework Tip Part 1 & 2 mandatory for Committed Listeners and MIT/Harvard students; Part 3 optional. Molecular weight — intact protein measurement. Primary amino acid sequence — peptide map. (Optional) Protein structure/shape — native vs denatured protein measurement.

Date: Wed Apr 23. Slides: Recitation Slides Recording: Recitation Recording

Tip This week’s practical focuses on LC‑MS workflows for eGFP characterization (intact mass and peptide mapping). On‑site work takes place at Waters Immerse Cambridge; remote students will use the shared dataset. Objective Connect measurement to design by acquiring and interpreting LC‑MS data. Concepts You’ll Learn Sample prep for intact mass vs peptide mapping Fundamentals of LC‑MS instrumentation & readouts Native vs denatured protein measurements Basic analysis and documentation

Immerse Cambridge (Waters) — About the lab Mass spectrometry overview (Proteomics) — Thermo Fisher Peptide mapping by LC/MS — Agilent (PDF) Intro to LC‑MS (lecture notes) — UCL (PDF)