Journal
ANALYTICAL CHEMISTRY
Volume 89, Issue 14, Pages 7302-7306Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b01848
Keywords
-
Categories
Funding
- National Institutes of Health [DK110665, GM110406]
Ask authors/readers for more resources
Equilibrium dialysis is a simple and effective technique used for investigating the binding of small molecules and ions to proteins. A three-dimensional (3D) printer was used to create a device capable of measuring binding constants between a protein and a small ion based on equilibrium dialysis. Specifically, the technology described here enables the user to customize an equilibrium dialysis device to fit their own experiments by choosing membranes of various material and molecular-weight cutoff values. The device has dimensions similar to that of a standard 96-well plate, thus being amenable to automated sample handlers and multichannel pipettes. The device consists of a printed base that hosts multiple windows containing a porous regenerated-cellulose membrane with a molecular-weight cutoff of similar to 3500 Da. A key step in the fabrication process is a print-pause-print approach for integrating membranes directly into the windows subsequently inserted into the base. The integrated membranes display no leaking upon placement into the base. After characterizing the system's requirements for reaching equilibrium, the device was used to successfully measure an equilibrium dissociation constant for Zn2+ and human serum albumin (K-d = (5.62 +/- 0.93) x 10(-7) M) under physiological conditions that is statistically equal to the constants reported in the literature.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available