Publisher Summary This chapter describes polyacrylamide gel electrophoresis of viral proteins. Viral structural and nonstructural proteins are agents of expression for the information carried in viral nucleic acid genomes. An understanding of the behavior of a virus requires its proteins be characterized. For this purpose and for the characterization of nucleic acids, electrophoresis in gels is the most sensitive and versatile method. The genetic content and number of proteins of most viruses is relatively small compared even to simple cells. Two aspects of gels as supporting media for zonal electrophoresis experiments are responsible for the dramatic separations. First, as simple anti-convection media gels, they are more homogeneous and stable than other commonly used materials. Second, with the correct choice of gel concentrations, the spaces in the gel matrix are of similar dimensions to macromolecules so that selective sieving occurs during the electrophoretic migration. Gels composed of polyacrylamide are extensively used at the present time.

Polyacrylamide Gel Electrophoresis of Viral Proteins

Polyacrylamide gel electrophoresis (PAGE) provides a versatile, gentle, high resolution method for fractionation and physical-chemical characterization of molecules on the basis of size, conformation, and net charge. The polymerization reaction can be rigorously controlled to provide uniform gels of reproducible, measurable pore size over a wide range. This makes it possible to obtain reproducible relative mobility (Rf) values as physical-chemical constants. Application and extension of Ogston9s (random fiber) model for a gel allows for calculation of molecular volume, surface area, or radius, free mobility, and valence from RJ measurements at several gel concentrations, to calculate gel concentration for optimal resolution, and to predict behavior of macromolecules on gel gradients by computerized methods. Extension of classical moving boundary theory has been used to generate multiphasic buffer systems (providing selective stacking, unstacking, restacking, and preparative steady-state-stacking) with known operating characteristics for any pH at 0° and 25°C. A general strategy for isolation of macromolecules and for macromolecular mapping has been developed. Preparative scale PAGE is operational for milligram loads and feasible for gram quantities.

https://science.sciencemag.org/content/sci/172/3982/440.full.pdf

Polyacrylamide Gel Electrophoresis

Estimation of molecular radius, free mobility, and valence using polyacrylamide gel electrophoresis☆

Free zone electrophoresis

Optimization in sample stacking for high-performance capillary electrophoresis

Some aspects of the use of "continuous" and "discontinuous" buffer systems in polyacrylamide gel electrophoresis.

Electrophoretic "particle sieving" in polyacrylamide gels as applied to ribosomes.

Apparatus for large-scale preparative polyacrylamide gel electrophoresis

Turnip yellow mosaic virus has been purified by electrophoretic “particle-sieving” in a polyacrylamide gel. This technique can be generally applied to fractionation of viruses, provided their shape and size allow them to penetrate the pores of the gel.

Sture Jerstedt

Papers

Some aspects of the use of "continuous" and "discontinuous" buffer systems in polyacrylamide gel electrophoresis.

Electrophoretic "particle sieving" in polyacrylamide gels as applied to ribosomes.

Apparatus for large-scale preparative polyacrylamide gel electrophoresis

“Particle-sieve” electrophoresis of viruses in polyacrylamide gels, exemplified by purification of turnip yellow mosaic virus