20.109(S20):Module 3

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20.109(S20): Laboratory Fundamentals of Biological Engineering

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Spring 2020 schedule        FYI        Assignments        Homework        Class data        Communication
       1. Screening ligand binding        2. Measuring gene expression        3. Engineering antibodies              


Module 3

Lecturer: Leslie McClain
Instructors: Noreen Lyell and Leslie McClain, and Becky Meyer
Research assistant: Sarah Cowles
TAs: Kevin Chung and Joe Kreitz
Lab manager: Hsinhwa Lee

Overview

An antibody is a soluble or membrane bound protein, produced by immune cells in blood. The purpose of an antibody is to recognize foreign substances, called antigens, and mark them for removal by cells that compromise our immune system. If one has a properly functioning immune system you've all been making new and improved antibodies your entire life.

Antibodies are also a critical reagent in medicine, research and diagnostics. Scientists have found ways to purify, modify and harvest antibodies from animals and to also engineer cells to produce specific antibodies. In Mod3 we'll use a method developed in the Wittrup lab to screen a yeast library of antibodies, against a particular antigen, using Fluorescence Assisted Cell Sorting, and try to identify a single antibody that shows improved antigen binding in a quantitative binding assay.

This module was developed thanks to the invaluable help and support of Wittrup lab PhD student, Sarah Cowles, and the generous contribution of reagents from Prof. Wittrup.

Schematic overview of the experimental approach in Module 3


Lab links: day by day

M3D1: Enrich candidate clones using fluorescence-activated cell sorter (FACS)
M3D2: Harvest candidate clones and prepare for sequencing
M3D3: Identify clones to characterize
M3D4: Characterize clone by titration using flow cytometry
M3D5: Analyze titration curves

Assignments

Research proposal presentations
Mini-report

References

Notes for teaching faculty

Prep notes for M3