PubMed · 6032232
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Y Nakayama, K Takeya. 1967-02-04. Esterase zymogram method for classifying mycobacteria.. https://doi.org/10.1038/213504a0
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Capillary electrophoresis was used for the analysis of recombinant human tumor necrosis factor beta (rhTNF-beta). Reproducible separation of the protein could be achieved in Polybrene-coated capillaries. Optimum conditions for separation were achieved with a pH 4.5 sodium acetate buffer. Besides Polybrene-coated capillaries, other commercially available column and column-coating reagents were also examined for the separation of rhTNF-beta. Capillary electrophoresis was found to be a potentially useful method for product analysis and process monitoring in recombinant-DNA technology.
A low-cost and highly sensitive laser-induced fluorescence detector for on-column detection systems was constructed and its applications were demonstrated in protein separation by capillary electrophoresis. The limit of detection (LOD) at a signal-to-noise ratio of 2 for Lissamine 20 was about 1.4 x 10(-21) mole at 10 nL sample injection and the relative standard deviation (RSD) for 1.5 x 10(-11) M of Lissamine 20 solution was about 15%. These values are comparable to or even better than those reported using a similar on-column detection system. This sensitive LIF detection was applied for the study of tryptic digestion of insulin and for urinary protein profiling.
Because the local electrical resistance is inversely proportional to the local cross-section of a capillary, the intensity of the electric field varies along the migration path if the inner diameter of the capillary is not constant. Therefore, fluctuations of the lateral dimensions of a capillary can directly affect the net elution time as well as the peak width, and, hence the final resolution. In this article, we develop the theoretical framework for the study of such effects. We then examine the simple case where both the mobility and the diffusion coefficient are field-independent; in particular, we demonstrate that resolution can be severely reduced if the inner walls are not flat, and that optimal resolution is always obtained for perfectly flat walls. Generalized to ultrathin gels, our results clearly indicate that both random and systematic variations of the gel thickness can greatly affect the performance of the separation process. Acceptable degrees of flatness are estimated for both geometries. This study thus provides a quantitative understanding of the type of quality control one requires to obtain optimal results with capillaries and ultrathin gels.