Premium
Rapid protein separation and diffusion coefficient measurement by frit inlet flow field‐flow fractionation
Author(s) -
Liu MinKuang,
Li Ping,
Giddings J. Calvin
Publication year - 1993
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560020917
Subject(s) - frit , chromatography , chemistry , fractionation , diffusion , analytical chemistry (journal) , inlet , flow (mathematics) , elution , channel (broadcasting) , volumetric flow rate , materials science , mechanics , thermodynamics , physics , computer science , computer network , mechanical engineering , engineering , metallurgy
In this study three flow field‐flow fractionation (flow FFF) channels are utilized for the separation of proteins and for the simultaneous measurement of their translational diffusion coefficients, D. One channel has a traditional sample inlet, whereas the other two incorporate a frit inlet design that permits more convenient and rapid sample introduction. The dependence of retention time on D , which leads to differential elution and the opportunity to measure D for protein peaks purified by the flow FFF process, is described theoretically and examined experimentally. Factors affecting band broadening, resolution, and optimization are also examined. The separation of proteins is achieved in the time range 4–20 min. Partial resolution is achieved in multiple runs requiring 2 min each. Values of D calculated from retention times are reported for 15 proteins. These include two protein dimers (bovine serum albumin and γ‐globulin) not ordinarily accessible to measurement. The D values from the three channels are compared with one another and with literature data. Reasonable consistency (within 3–4%) is found. High‐speed repetitive runs can be used to acquire multiple values of D in time intervals as short as 1 min.