Ross Kieser,  Braxton Bliss, Ben Feikema, Zac Ardavanis, Jose Abraham Mena

Our Focus

Protein crystallization is an important tool in understanding macromolecules’ structures; determining a macromolecule’s structure is crucial for the efficient development of drugs and treatments that target and interact with it. Protein crystals are produced when the solution the protein is dissolved in becomes supersaturated. Several common methods including vapor diffusion, microbatch, and microdialysis. Among the major methods, vapor diffusion is the most used in industry and scientific research. Vapor diffusion accomplishes supersaturation using concentration gradients as the driving force. 

Protein crystallization experiments are hard to generalize because each molecule behaves differently under different conditions; therefore, optimization is based on a per-case basis. Take lysozyme for instance; the conditions under which crystallization of lysozyme occurs have been optimized to the point that commercial kits are available. The variables that affect the rate of crystallization include pH, relative humidity, protein concentration, solvent, presence and concentration of precipitating agent and/or salts, and temperature, among others. 

This design project aims at investigating the effects of relative humidity in a hanging drop vapor diffusion context with the end goal of developing an optimized technique for the crystallization of a protein of interest. 

What We Accomplished

Crystallization with Hanging Drop Vapor Diffusion
Traditional Humidity Control Solvent Used in Column
Lysozyme saturated water
Bovine Serum Album (BSA) X tris-HCL

Our Humidity Control Apparatus

Displays the column used to provide saturated air with desired solvent to the crystallization tray. Air enters from the right while saturated air comes from the top.
Displays the delivery system for adding saturated air to each well. Saturated air enters into the control volume below and is introduced to each well by a dispensing needle. One needle goes from the control volume to the well, whereas the other goes from the well to the environment.

 

 

 

 

 

    

Our Crystals

Lysozyme crystals.
BSA crystals.

Check Out Our Project Proposal and Brief

Team 21 PPFS 

Project Brief