A new two-dimensional gel electrophoresis system for the analysis of complex protein mixtures: Application to the ribosome of E. coli

1975 ◽  
Vol 2 (1) ◽  
pp. 35-40 ◽  
Author(s):  
U. C. Knopf ◽  
A. Sommer ◽  
J. Kenny ◽  
R. R. Traut
Keyword(s):  
Gel Electrophoresis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
E Coli ◽  
Complex Protein ◽  
Electrophoresis System

scholarly journals Combination of strong anion exchange liquid chromatography with microchip capillary electrophoresis sodium dodecyl sulfate for rapid two-dimensional separations of complex protein mixtures

2021 ◽  
Author(s):  
Holger Zagst ◽  
Christin Elgert ◽  
Sönke Behrends ◽  
Hermann Wätzig
Keyword(s):  
Capillary Electrophoresis ◽  
Liquid Chromatography ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Soluble Guanylyl Cyclase ◽  
Two Dimensional ◽  
Microchip Capillary Electrophoresis ◽  
Two Dimensional Gel Electrophoresis ◽  
Complex Protein ◽  
Strong Anion Exchange

AbstractTwo-dimensional separations provide a simple way to increase the resolution and peak capacity of complex protein separations. The feasibility of a recently developed instrumental approach for two-dimensional separations of proteins was evaluated. The approach is based on the general principle of two-dimensional gel electrophoresis. In the first dimension, semi-preparative strong anion exchange high-performance liquid chromatography is utilized and fractions are collected by means of a fraction collector. They are subsequently analyzed in the second dimension with microchip capillary electrophoresis sodium dodecyl sulfate. Microchip capillary electrophoresis provides the necessary speed (approximately 1 min/fraction) for short analysis. In this study, three different samples were investigated. Different constructs of soluble guanylyl cyclase were expressed in Sf9-cells using the baculovirus expression system. Cell lysates were analyzed and the resulting separations were compared. In our experimental setup, the soluble guanylyl cyclase was identified among hundreds of other proteins in these cell lysates, indicating its potential for screening, process control, or analysis. The results were validated by immunoblotting. Samples from Chinese hamster ovary cell culture before and after a purification step were investigated and approximately 9% less impurities could be observed. The separation patterns obtained for human plasma are closely similar to patterns obtained with two-dimensional gel electrophoresis and a total of 218 peaks could be observed. Overall, the approach was well applicable to all samples and, based on these results, further directions for improvements were identified. Graphical abstract .

Evaluation of antibody drugs using automated two dimensional gel electrophoresis system

2018 ◽  
Vol 62 (1) ◽  
pp. 13-18
Author(s):  
Hideki Kinoshita ◽  
Masayuki Kosugi ◽  
Kimihiko Yabe ◽  
Takateru Matsunaga ◽  
Mitsuhiro Kinoshita
Keyword(s):  
Gel Electrophoresis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Electrophoresis System

Two-dimensional gel electrophoresis of serum specimens from patients with monoclonal gammopathies.

1982 ◽  
Vol 28 (4) ◽  
pp. 900-907 ◽  
Author(s):  
R P Tracy ◽  
R M Currie ◽  
R A Kyle ◽  
D S Young
Keyword(s):  
Gel Electrophoresis ◽  
Light Chains ◽  
Correct Identification ◽  
Two Dimensional ◽  
Excellent Correlation ◽  
Serum Samples ◽  
Two Dimensional Gel Electrophoresis ◽  
Monoclonal Gammopathies ◽  
Heavy Chains ◽  
Electrophoresis System

Abstract We modified the ISO-DALT two-dimensional gel electrophoresis system to allow the routine examination of serum specimens from patients with monoclonal gammopathies. This system, MC-Iso 1, is characterized by a broad pH gradient for resolving the basic immunoglobulin heavy and light chains. The increased resolution of basic proteins may be explained on theoretical grounds by an increase in voltage in this region of the cell. Ancillary techniques, such as those for albumin removal and pI assignment through use of charge standards, have also been implemented. The locations of immunoglobulin heavy chains have been confirmed by examination of over 250 serum samples as well as by "electro-blotting," with use of specific antisera. IgG subclass may also be predicted by location, but not with perfect accuracy. Differentiation of kappa and lambda light chains by relative mobility has been examined; the predictive value for correct identification of kappa chains is 83%, that for lambda chains 69%. Several unknown proteins have been observed in macroglobulinemia, related to mu heavy chain. Finally, we have determined that there is excellent correlation between non-denaturing isoelectric focusing and our system for pI assignment of light chains. This has importance due to reports of the potential importance of light-chain pI in the development of renal disease in patients with monoclonal gammopathies.

Sign in / Sign up

Export Citation Format

Share Document