FPLC—Fast Protein Liquid Chromatography

Gel Filtration by Fast Protein Liquid Chromatography (FPLC)... 

P Barrefors, B Ekstrand, L Fagerstam, M Larsson-Raznikiewicz, J Schaar, P Steffner. Fast protein liquid chromatography (FPLC) of bovine caseins. Milchwissenschaft 40 257-260, 1985. 

High-performance liquid chromatography (HPLC) and fast protein liquid chromatography (FPLC) rely on the same separation principles as the traditional chromatography columns, but tend to be much faster because of high flow rates that are possible due to the uniform bead size and the mechanical strength of the beads. See also Chapter 4, section 1.2.2. 

Fast protein liquid chromatography (FPLC) equipment. 

Fig. 6. Comparison of immunoassays using clonotype antibodies. The EDso values (thyroxine concentration corresponding to 50% of bound form at zero concentration of thyroxine) in thyroxine immunoassays using clonotype antithyroxine antibody prepared by fast protein liquid chromatography (FPLC). RIA, double-antibody radioimmunoassay FLA, fluorescence polarization immunoassay EIA, enzyme immunoassay. [Cited from Miyai et al. (M5).]...

There are several problems requiring careful attention. Lysozyme has a tendency to form complexes with many substances [e.g., alkyl sulfates, fatty acids, aliphatic alcohols (Smith and Stocker, 1949), cephalins (Brusca and Patrono, 1960), and other proteins]. Of particular importance is its tendency to form complexes with transferrins [e.g., ovotrans-ferrin (Ehrenpreis and Warner, 1956)]. These interactions lead to difficulties in the isolation of lysozyme. Some recent workers have used fast protein liquid chromatography (FPLC) and high-performance liquid chromatography (HPLC) (e.g., Ekstrand and Bjorck, 1986). The resolution in these procedures may not always be satisfactory, and in HPLC pressure and solvent effects must be monitored carefully if the product is to be suitable for conformation and activity studies. 

Triethanolamine buffer 0.02M triethanolamine, pH 7.7. For ionic gradient separations by fast-protein liquid chromatography (FPLC), also use triethanolamine buffer containing IMNaCl. Store at 4°C. 

Another example of on-line monitoring of enzyme activities was given by Kunnecke et al. [88], when a FIA-system was used for the determination of enzyme activities during protein purification by fast protein liquid chromatography (FPLC). Photometric assays for four different oxidases were established in a FIA-system extending the linear range by the so-called zone sampling method. The FIA-device was coupled to the FPLC unit behind a... 

Svete, P. Milacic, R. Mitrovic, B. Pihlar, B. Potential for the speciation of Zn using fast protein liquid chromatography (FPLC) and convective interaction media (CIM ) fast monolithic chromatography with FAAS and electrospray (ES)-MS-MS detection. Analyst 2001, 126, 1346-1354. 

Determination of the radiochemical purity using fast protein liquid chromatography (FPLC) and instant thin layer chromatography (ITLC), and purification using the SepPak procedure and size exclusion chromatography. 

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