Cerebrospinal fluid protein electrophoresis

PI. Pirttila, T., Frey, H., Mattila, K., and Siden, L., Immobilized pH gradient isoelectric focusing of cerebrospinal fluid proteins. Electrophoresis 9, 582-588 (1988). 

W5. Windisch, R. M., and Bracken, M. M., Cerebrospinal fluid proteins Concentration by membrane ultrafiltration and fractionation by electrophoresis on cellulose acetate. Clin. Cherru 16,416-419 (1970). 

Oligoclonal bands Two to five bands of immunoglobulins on protein electrophoresis of cerebrospinal fluid. 

Many biological fluids (e.g., serum and plasma) contain sufficient protein without excessive salt concentration to be applied at once to the paper. Some other fluids, such as urine, may need preliminary dialysis because they contain too much salt, and they often need concentration also. Still other fluids, such as cerebrospinal fluid, always need concentration before electrophoresis. 

Davidsson, P., Westman, A., Puchades, M., Nilsson, C.L., and Blennow, K. (1999). Characterization of proteins from human cerebrospinal fluid by a combination of preparative two-dimensional liquid-phase electrophoresis and matrix-assisted laser desorption/ioniza-tion time-of-flight mass spectrometry. Anal. Chem. 71, 642-647. 

Most proteomics analyses involve 2-D electrophoresis and MALDl-TOF MS steps, as this approach allows for a reliable quantification of changes in protein levels. In one study, the LC-MS/MS approach was used for the detection of proteins enriched in amyloid plaques in the AD brain (liao et al., 2004). Employing the former proteomic approach, levels of about 100 proteins were foimd to be changed in the AD brain and cerebrospinal fluid (CSF) in comparison with the control brain and CSF, respectively (Table 15.1). These proteins have various functions, mainly involved in neurotransmission, guidance, signal transduction, metabolism, detoxification, and conformational changes. The CSF proteins are plasma proteins. 

Even though free-solution CE is most commonly used for neuropeptides and neuroproteins, other forms of CE have also been employed. For instance, as an alternative to conventional slab-gel electrophoresis, a method using sodium dodecyl sulfate (SDS) capillary gel electrophoresis was developed. It was applied to low-molecular-mass proteins (j8-trace protein, ft-microglobulin, -trace protein, and myelin basic protein) in cerebrospinal fluid [4], Advantageous features of capillary gel electrophoresis over slab-gel electrophoresis are compatibihty with small sample volumes, shorter analysis times, and more accurate quantihcation of the analytes. 

After 10 days of antibiotic treatment, the patient was afebrile and cerebrospinal fluid was sterile, with normal protein and glucose levels. Drug treatment was discontinued, but after 2 days. she developed vomiting and fever to 40.5 °C. Cerebrospinal fluid culture was sterile, but counterimmuno-electrophoresis (CIE) was po.sitivefor H influenzae type b polyribosylribitol phosphate antigen. 

Wait, R., Gianazza, E., Eberini, I., Sironi, L., Dunn, M.J., Gemeiner, M., and Miller, I., 2001, Proteins of rat serum, urine, and cerebrospinal fluid VI. Further protein identifications and interstrain comparison. Electrophoresis 22 3043-3052. 

Hiroaka, A. et al.. Charge microheterogeneity of the fi-trace proteins (lipocalin-type prostaglandin D synthase) in the cerebrospinal fluid of patients with neurological disorders analyzed by capillary isoelectric focusing. Electrophoresis, 22, 3433, 2001. 

Unlike mass spectroscopy, gel electrophoresis does not provide a quantitative value for the amount of given protein. However, it provides a low cost and relatively rapid method for the analysis of multiple proteins in a specimen, especially when implemented as a capillary electrophoresis system. Therefore, it has been used for the separation of enzymes (e.g., creatinine phosphokinase), mucopolysaccharides, plasma, serum, cerebrospinal fluid, urine, and other bodily fluids [13]. It is also used for quality control applications for the manufacturing of biological compounds to verify the purity or to examine the manufacturing yield [14]. 

The main clinical significance of cerebrospinal fluid (CSF) protein electrophoresis is for the detection of the oligoclonal bands, which are present in multiple sclerosis in the gamma region. Because proteins in CSF occur at much lower concentrations than in serum (100 times less), a 10-20-fold sample concentration is needed. CSF protein separation can be accomplished in less than 10 min with CE versus 2h for AG with the ability to detect oligoclonal banding by this technique. 

Hiraoka, A. Arato, T. Tominaga, I. Eguchi, N. Oda, H. Urade, Y. Analysis of low-molecular-mass proteins in cerebrospinal fluid hy sodium dodecyl sulfate capillary gel electrophoresis. J. Chromatogr. B, 1997, 697,141 147. 

---