» Articles » PMID: 38301115

Versatile Capillary Cells for Handling Concentrated Samples in Analytical Ultracentrifugation

Overview
Journal Anal Chem
Specialty Chemistry
Date 2024 Feb 1
PMID 38301115
Authors
Affiliations
Soon will be listed here.
Abstract

In concentrated macromolecular dispersions, far-from-ideal intermolecular interactions determine the dispersion behaviors including phase transition, crystallization, and liquid-liquid phase separation. Here, we present a novel versatile capillary-cell design for analytical ultracentrifugation-sedimentation equilibrium (AUC-SE), ideal for studying samples at high concentrations. Current setups for such studies are difficult and unreliable to handle, leading to a low experimental success rate. The design presented here is easy to use, robust, and reusable for samples in both aqueous and organic solvents while requiring no special tools or chemical modification of AUC cells. The key and unique feature is the fabrication of liquid reservoirs directly on the bottom window of AUC cells, which can be easily realized by laser ablation or mechanical drilling. The channel length and optical path length are therefore tunable. The success rate for assembling this new cell is close to 100%. We demonstrate the practicality of this cell by studying: (1) the equation of state and second virial coefficients of concentrated gold nanoparticle dispersions in water and bovine serum albumin (BSA) as well as lysozyme solution in aqueous buffers, (2) the gelation phase transition of DNA and BSA solutions, and (3) liquid-liquid phase separation of concentrated BSA/polyethylene glycol (PEG) droplets.

Citing Articles

Amino acids modulate liquid-liquid phase separation in vitro and in vivo by regulating protein-protein interactions.

Xu X, Rebane A, Roset Julia L, Rosowski K, Dufresne E, Stellacci F Proc Natl Acad Sci U S A. 2024; 121(50):e2407633121.

PMID: 39642205 PMC: 11648668. DOI: 10.1073/pnas.2407633121.

References
1.
Minton A, Lewis M . Self-association in highly concentrated solutions of myoglobin: a novel analysis of sedimentation equilibrium of highly nonideal solutions. Biophys Chem. 1981; 14(4):317-24. DOI: 10.1016/0301-4622(81)85033-8. View

2.
Xu X, de With G, Colfen H . Self-association and gel formation during sedimentation of like-charged colloids. Mater Horiz. 2022; 9(4):1216-1221. DOI: 10.1039/d1mh01854j. View

3.
To S, Brautigam C, Chaturvedi S, Bollard M, Krynitsky J, Kakareka J . Enhanced Sample Handling for Analytical Ultracentrifugation with 3D-Printed Centerpieces. Anal Chem. 2019; 91(9):5866-5873. PMC: 6750210. DOI: 10.1021/acs.analchem.9b00202. View

4.
Chao Y, Shum H . Emerging aqueous two-phase systems: from fundamentals of interfaces to biomedical applications. Chem Soc Rev. 2019; 49(1):114-142. DOI: 10.1039/c9cs00466a. View

5.
Brian A, Frisch H, LERMAN L . Thermodynamics and equilibrium sedimentation analysis of the close approach of DNA molecules and a molecular ordering transition. Biopolymers. 1981; 20(6):1305-28. DOI: 10.1002/bip.1981.360200615. View