Protein separation techniques

FIGURE 10.1 Schematic overview of protein separation techniques. 

Improved protein separation techniques utilizing hquid chromatography and electrophoresis coupled with X-ray diffraction and NMR studies have given insights into the three-dimensional structures of cellulolytic enzymes. This molecular architecture data coupled with DNA sequence information has given clues to the chemical mechanisms of enzymatic hydrolysis and molecular interaction between cellulose and the enzymes. 

Pungor, E. Afeyan, N. B. Gordon, N. F. Cooney, C. L. "Continuous Affinity-recycle Extraction A Novel Protein Separation Technique" BiojTechnology. 1987,5, pp 604-608. 

The last decade has seen the development of several high-resolution protein separation techniques and three of these—isotachophoresis, high-performance liquid chromatography, and lEF/SDS gradient PAGE (ISO-DALT)—have been discussed in this review. The three techniques complement each other in both the method of separation and the method of detection. 

Development of electrophoretic protein separation techniques have been paralleled by improvements in protein detection methods. Protein detection in early electrophoretic applications, utilizing electrophoretic separations of solutions or colloidal suspensions from about 1816 to 1937, was limited to direct visualization of proteins coated onto microspheres, or studies of naturally colored proteins such as hemoglobin, myoglobin, or ferritin <1-4). An increase in sensitivity and the ability to detect non-colored proteins was achieved by the use of the specific absorption, by proteins, of ultraviolet light. This detection technique permitted Tiselius,in 1937, to demonstrate the quantitative electrophoretic separation of ovalbumin, serum globulin fractions and Bence Jones proteins (S). Tiselius also employed the shadows, or schlieren, created by the boundaries, due to the different concentrations of proteins in the electrophoretic system to detect protein position and concentration ( ). These detection methods served as the main methods for protein detection in the liquid electrophoresis systems. However,... 

Recent developments In biology have ushered In a "revolution" In our knowledge of the cell, particularly the microbial cell. For the familiar bacillus Escherichia coll 3/5 to 4/5 of all Its molecular elements are known and new protein separation techniques are speeding this Identification process to a conclusion. Thus, we shall soon have the complete catalogue of E. coll parts. However, we still know relatively little about the Integrated behavior of the cell, how It will behave when Its molecular elements are changed or en It finds Itself In a novel environment. 

The properties are very special in that the proteins attach to more acidic proteins. Before modern protein separation techniques became available, the protamines were used for purification of other proteins (Green and Hughes, 1955). The tight binding to other proteins was also utilised, and still is, for the introduction of insulin to diabetic patients (Hagedorn et al., 1936). It was in fact the protamine from salmon (salmin) that made insulin injection a success, in that it allowed for a slow release of insulin into the blood. This had previously been a problem. 

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