Preparative isoelectric focusing in a cellulose‐based separation medium

• Published in: Journal of separation science Vol. 40; no. 11; pp. 2498 - 2505
• Main Authors: Salplachta, Jirí, Horka, Marie, Slais, Karel
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01.06.2017
• Subjects: Analytical chemistry; Buffers; Cellulose; colored pI markers; Cytochromes c - isolation & purification; Electrophoresis; Ethylene glycol; Evaporation; Genotype & phenotype; Isoelectric Focusing; Markers; Mathematical analysis; Mathematical models; preparative; Proteins; Recovery; Segments; Separation; Serum albumin; Serum Albumin, Bovine - isolation & purification; separation; isoelectric focusing; proteins; colored pI markers; preparative
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.201700036

An improved preparative method based on isoelectric focusing of analytes in a cellulose‐based separation medium is described in this study. Cellulose is suspended in an aqueous solution of simple buffers, ethylene glycol, glycerol, nonionic surfactant, and colored pI markers. Water partially evaporates during focusing run and the separation takes place in an in situ generated layer of cellulose, which has a gel‐like appearance at the end of analysis. Final positions of analytes are indicated by the positions of zones of focused pI markers. Fractions, segments of the separation medium with analytes, can be simply collected by spatula and analyzed by downstream analytical methods. Good focusing ability of the new method and almost quantitative recovery of model proteins, cytochrome c and bovine serum albumin, was verified by gel electrophoresis and capillary isoelectric focusing of the collected fractions.