Asymmetric flow Field-Flow Fractionation

The asymmetric flow Field-Flow Fractionation (AsFlFFF) system, is coupled with photodiode array UV/Vis detector, fluorescence detector and refractive index detectors (two mass detectors), and multiangle static and dynamic light scattering detectors.

The new light scattering (LS) detector (DAWN HELEOS II) and refractive index (RI) detectors (OptiLab rEX) have ~60 times bigger dynamic range, which allows determination of MWs of protein samples from ~ 1 ug/ml to a few mg/mL (concentration used during structural studies by NMR or X-ray).

I may also include fluorescence detection for analysis of eluting peaks- please indicate such need if your research may benefit from this feature.

We plan on installing an automated gradient set up (as described by Attri and Minton, (2005a, 2005b) to analyze protein association processes. Such gradient experiment (either with one protein or mixing two interacting partners) would allow determination of association constant.

First time users of the resource follow the booking and charging protocol; please fill out the Biophysics Resource Usage Form, and email the completely filled usage form, with email Subject: "Biophysics Resource Usage," to Ewa Folta-Stogniew.

**Attention All Mac users using "Preview" instead of "Acrobat", please save PDFs as follows: 1. Under the "File" pulldown menu, Select "Print…" 2. Click "PDF" button in lower left hand corner of window. 3. In the pulldown menu Select "Save As PDF." 4. Save PDF.**


The FFF technology is reviewed in:

Giddings, JC (1993) Science, 260, 1456-1466
"Field-flow fractionation: Analysis of macromolecular, colloidal, and particulate materials"

Fraunhofer, W; Winter, G (2004) European Journal of Pharmaceutics and Biopharmaceutics, 58, 369-383
"The use of asymmetrical flow field-flow fractionation in pharmaceutics and biopharmaceutics"

Reschiglian, Pierluigi; Zattoni, Andrea; Roda, Barbara; Michelini, Elisa; Roda, Aldo (2005) Trends in Biotechnology, 23, 475-483
"Field-flow fractionation and biotechnology"

Selected Applications:

Nanoparticles as drug carriers
Fraunhofer, W; Winter, G; Coester, C (2004) Anal Chemistry, 76, 1909-1920
"Asymmetrical Flow Field-Flow Fractionation and Multiangle Light Scattering for Analysis of Gelatin Nanoparticle Drug Carrier Systems"

Prion proteins:
Silveira, Jay R; Raymond, Gregory J; Hughson, Andrew G; Race, Richard E; Sim, Valerie L; Hayes, Stanley F; Caughey, Byron (2005) Nature 437, 257-261 (8 September 2005)
"The most infectious prion protein particles"

Liposomes:
Shuliang Li and Andrew F. Palmer (2004) Langmuir, 20, 4629-4639
"Effect of Actin Concentration on the Structure of Actin-Containing Liposomes"

Korgel, Brian A; van Zanten, John H; Monbouquette, Harold G (1998) Biophys. J., 74, 3264-3272
"Vesicle Size Distributions Measured by Flow Field-Flow Fractionation Coupled with Multiangle Light Scattering"

Moon, MH; Park, I; Kim, Y (1998)  J. Chromatography A, 813, 91-100
"Size characterization of liposomes by flow field-flow fractionation and photon correlation spectroscopy. Effect of ionic strength and pH of carrier solutions"

Ribosomes:
Andersson, CI; Arfvidsson, C; Kallio, PT; Wahlund, KG; Bulow, L (2003) Biotechnol. Lett. 25, 1499-1504
"Enhanced ribosome and tRNA contents in Escherichia coli expressing a truncated Vitreoscilla hemoglobin mutant analyzed by flow field-flow fractionation"

Environmental samples:
Baalousha, M; Kammer, FVD; Motelica-Heino, M; Le Coustumer, P (2005) Journal of Chromatography A, 1093, 156-166
"Natural sample fractionation by FlFFF-MALLS-TEM: Sample stabilization, preparation, pre-concentration and fractionation"