Cerebrospinal fluid proteins analysis

Sjogren, M., Blennow, K. (2002). Proteome analysis of cerebrospinal fluid proteins in Alzheimer patients. Neuroreport 16, 611-615. 

Kastenbauer S, Angele B, Sporer B, Pfister HW, Koedel U (2005) Patterns of protein expression in infectious meningitis A cerebrospinal fluid protein ai ray analysis. J Neuroimmunol 164(1-2) 134-139. 

The determination of ammonia after the regular or modified Kjeldahl digestion presents rather less serious problems than those already dis cussed. The advantages of the micro-Kjeldahl distillation (69, 80, 81, 82, 83) as compared with the macro>method, or even the semimicro-method are now generally recognized. A comparative study of the macro-and microscale determination in the analysis of flour, wheat and com for their protein content was made by Robinson and Shellenberger (27). The micro-Kjeldahl method has been used for systematic plasma protein analysis (84, 85), saliva proteins (86), milk proteins (87), and cerebrospinal fluid protein (88),... 

A generalized systemic illness may accompany HIV seroconversion (Cooper et al. 1985). Guillain-Barre syndrome (GBS) (Piette et al. 1986), unilateral (Wiselka et al. 1987) or bilateral facial palsies (Wechsler and Ho 1989), bibra-chial palsy (Calabrese et al. 1987) and sensory neuropathy (Denning 1988) have been reported to occur during this process, usually within 1-2 weeks of the acute febrile illness. Spinal fluid analysis may show a mild to moderate mononuclear pleocytosis and a mild increase in protein levels. The precise relationship to HIV viral load in the cerebrospinal fluid (CSF) or plasma is unknown (Brew 2003). There is no proven therapy, but most patients recover spontaneously without any treatment. 

The use of amino acid analysis to characterize protein hydrolyzates is well known, and IEC is an integral part of the commercial amino acid analyzer.61 Determinabon of individual amino acids in physiological fluids such as plasma and cerebrospinal fluid can be important in the etiology of various diseases.62 63 Of particular current interest is the measurement of total... 

Hiraoka, A., et al (1998). Sodium dodecyl sulfate-capillary gel electrophoretic analysis of molecular mass microheterogeneity of beta-trace protein in cerebrospinal fluid from patients with central nervous system diseases./. Chromatogr. A 802, 143-8. 

A. Westman, C. L. Nilsson, and R. Ekman. Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Analysis of Proteins in Human Cerebrospinal Fluid. Rapid Commun. Mass Spectrom., 12(1998) 1092-1098. 

This technique is based on the complexity of the system that is not favored in proteome analysis. It is preferred in the separation of some proteins in biological fluids (e.g., cerebrospinal fluids) and protein-protein interactions where relatively few proteins are involved. 

Ogata Y Charlesworth MC, Muddiman DC (2005) Evaluadon of protein depledon methods for the analysis of total-, phospho-and glycoproteins in lumbar cerebrospinal fluid. J Proteome Res 4 837-845. 

Yuan X, Desiderio DM (2003) Proteomics analysis of phosphotyrosyl-proteins in human lumbar cerebrospinal fluid. J Proteome Res 2 476 87. 

The bioanalyst can be required to analyse most biofluids although the most common are urine and the aqueous phase of blood, i.e. plasma or serum. Other samples may be cell and tissue extracts, synovial fluid, cerebrospinal fluid (CSF) and saliva. In the case of urine and CSF with their very low protein content it might be possible to directly inject the sample into an HPLC column. With most silica-based packing materials, direct injection of blood proteins will rapidly lead to column deterioration. HPLC columns are expensive and their efficiency is easily lost so correct preparation of samples will not only improve column life but also improve the results. At its simplest it is only necessary to remove particulate matter from samples to prevent clogging of the column and frits. Modern HPLC packings are very susceptible to contamination by proteins, fats and other macromolecules from biological samples and it is necessary to remove these (except of course for protein analysis). 

Determination of these compounds is carried out frequently in biological fluids. Analysis in urine requires a previous filtration to remove cells and other particulate matter then, the samples are diluted and directly injected onto the column. With cerebrospinal fluid, the samples are obtained by lumbar puncture each ahquot is centrifuged and decanted before analysis. Often in plasma or semm, some form of protein removal is needed because the presence of these compounds in injected samples can cause modifications of the column and bias in chromatographic results. Protein removal can be performed by various methods such as protein precipitation, ultrafiltra-tion, centrifugation, liquid-phase or solid-phase extraction, and column-switching techniques. 

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. 

Paper chromatographic methods have been devised to determine cephalothin and its micro-biologically active metabolite deacetylcephalothin in body fluids. Miller developed a method that is satisfactory for analysis of urine samples. Hoehn e t al. odescribed a method that affords quantitative disassocia-tion of cephalothin from plasma proteins, and developed a chromatographic technique to measure low levels of cephalothin and its metabolite in urine, plasma, synovial fluid, and cerebrospinal fluid. 

Protein content, particularly in urine or cerebrospinal fluid, may also be estimated by methods based on precipitation using sulfosalicylic acid (an anionic protein precipitant) or heat. The turbidity, which is a measure of protein concentration, can be quantitated by spectropho-tometric absorbance methods or light scattering analysis. Absorbance of a hydrophobic indicator dye that binds to protein and changes color is also used. 

Cerebrospinal fluid. Both PAGE/gradient PAGE and lEF/gradient PAGE have been used in the analysis of csf proteins (44-73 components... 

Lumbar puncture and cerebrospinal fluid examination Paracentesis Pericardiocentesis Secretin-pancreozymin Semen analysis Sims-Huhner test Sweat electrolytes test Tau protein... 

There are a range of applications of conventional cIEF to real-world analysis, including the characterization of protein samples in laboratories of biotech pharmaceutical companies, and clinical analysis. In the pharmaceutical industry, some methods have been developed based on this technology [13,14] and validated [15-17] as the identity assays in regulated pharmaceutical laboratories for the analysis of therapeutic monoclonal antibodies and glycoproteins. cIEF has also been used in clinical analysis for human hemoglobin analysis [18,19] and the analysis of proteins in cerebrospinal fluid [20]. There are several reviews on the applications of the conventional cIEE [10,21]. However, the pace of acceptance of conventional cIEF for the analysis of real-world samples is slow. 

The main clinical significance of cerebrospinal fluid (CSF) protein eleetrophoresis is for the detection of the oligoclonal bands, which are present in multiple sclerosis in the gamma region. Similar to urine, proteins in the CSF are present fluid in very low concentration (100 times less than serum). For the majority of the samples, a 10- to 20-fold concentration is preferred before analysis by CE (by the same membrane concentrators used for urine). CSF protein separation can be accomplished in less than 10 min with CE versus 2 h for AG with the ability to detect oligoclonal banding by this technique [36]. 

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