field flow fractionation in biopolymer analysis

[...]... Time-minimized determination of ribosome and tRNA levels in bacterial cells using flow field-flow fractionation Anal Biochem 313(1):76–85 Chapter 2 Assessing Protein-Ultrafiltration Membrane Interactions Using Flow Field- Flow Fractionation Galina E Kassalainen and S Kim Ratanathanawongs Williams Abstract Flow FFF (FlFFF) is used to rapidly and conveniently measure initial stage protein fouling on ultrafiltration... 1 ml; HSA, 1.25 mg/ml, 9 ml Sample loading: flowrate ¼ 0.1 ml/min, loop volume ¼ 10 ml, time ¼ 1 min Relaxation/focusing: focusing point (distance from inlet, z´) ¼ 5.0 cm, Fcross ¼ 5 ml/min during 1 min and 9.9 ml/min during 15 s Elution: Fin ¼ 9.7, Fcross ¼ 8.9, Fout ¼ 0.8 ml/min, to ¼ 0.09 min (Reproduced from [14], # 1989, with permission from Elsevier) 1 Flow FFF – Basics and Key Applications 15...x Contents 8 Field- Flow Fractionation for Assessing Biomolecular Interactions in Solution 113 Robert Y -T Chou, Joey Pollastrini, Thomas M Dillon, Pavel V Bondarenko, Lei-Ting T Tam, Jill Miller, Michael Moxness, and Shawn Cao 9 Flow Field- Flow Fractionation: Analysis of Biomolecules and Their Complexes ... multi-angle light scattering Starch-Starke 58(2):55–65 44 Wahlund K-G (2000) Asymmetrical flow field-flow fractionation In: Schimpf M, Caldwell K, Giddings JC (eds) Field- flow fractionation handbook Wiley-Interscience, New York, pp 279–294 45 Schimpf ME (2000) Resolution and fractionating power In: Schimpf M, Caldwell K, Giddings JC (eds) Field- flow fractionation handbook Wiley-Interscience, New York, pp... Fcross/Fout constant This is simply effected by increasing Fin as much as possible If analysis time can be sacrificed, leading to higher retention times, a second approach is to increase Fcross at constant Fin by increasing the ratio Fcross/Fout, i.e by decreasing Fout For some instruments there are upper limitations in the available crossflow rates due to the pumping system and/or flow regulators This can... Mines, Golden, CO, USA e-mail: krwillia@mines.edu S.K.R Williams and K.D Caldwell (eds.), Field- Flow Fractionation in Biopolymer Analysis, DOI 10.1007/978-3-7091-0154-4_2, # Springer-Verlag/Wien 2012 23 24 2.1 G.E Kassalainen and S.K.R Williams Introduction Flow field-flow fractionation (FlFFF) has become the most widely used technique of the FFF family [1–5] Reasons include the need for a low shear rate... protein separation and characterization Anal Biochem 81(2):395–407 1 Flow FFF – Basics and Key Applications 19 6 Giddings JC, Lin G-C, Myers MN (1978) Fractionation and size distribution of water soluble polymers by flow field-flow fractionation J Liq Chromatogr 1(1):1–20 7 Giddings JC, Lin G-C, Myers MN (1978) Fractionation and size analysis of colloidal silica by flow field-flow fractionation J Colloid Interf... particles in asymmetrical flow field-flow fractionation Flow conditions for rapid elution Anal Chem 74(21):5621–5628 34 Giddings JC (1973) Conceptual basis of field-flow fractionation J Chem Educ 50:667 35 Giddings JC (1993) Field- flow fractionation: analysis of macromolecular, colloidal, and particulate materials Science 260:1456–1465 36 Ratanathanawongs-Williams SK (2000) Flow field-flow fractionation In: Schimpf... molecular size which indicates changes in structural distributions within the biopolymer [48–52] 1.5 Key Applications Below are reported some pioneering and ground-breaking studies using AsFlFFF 1.5.1 Proteins – Covalent/Non-covalent Aggregates, Antibody Aggregates Some of the earliest examples of the power of AsFlFFF as a fractionation technique for biopolymers were fractionation of proteins and aggregates... ultra-high molar mass linear biopolymers care must be taken to avoid mass overloading by chain entanglement and shear degradation For much smaller polynucleotides such as tRNA the flow FFF experiments are straight-forward since mass overloading does not occur easily This was utilized in determinations of tRNA and ribosome levels in genetically engineered bacterial cells [64] by injecting the cell lysate . class="bi x0 y0 w0 h0" alt="" Field-Flow Fractionation in Biopolymer Analysis . S. Kim R. Williams l Karin D. Caldwell Editors Field-Flow Fractionation in Biopolymer Analysis Editors Prof. S. Kim. Springer-Verlag/Wien SpringerWienNewYork is a part of Springer Science+Business Media springer.at Typesetting: SPi, Pondicherry, India Printed on acid-free and chlorine-free bleached paper SPIN:. Assessing Protein-Ultra filtration Membrane Interactions Using Flow Field-Flow Fractionation 23 Galina E. Kassalainen and S. Kim Ratanathanawongs Williams 3 Hollow-Fiber Flow Field-Flow Fractionation: