Proteinas do liquido cefalorraqueano: I. Estudo comparativo entre metodos de concentração

Vermes, Lucia M. Singer

doi:10.1590/S0004-282X1983000100001

Resumos

No intuito de escolher entre métodos de concentração, aplicáveis ao LCR, aquele que, não provocando desnaturação de proteínas, seja o mais exato, preciso e de fácil aplicação em laboratórios de rotina, foram estudados quatro procedimentos: diálise contra solução de goma-arábica, centrífugo-ultrafiltração, absorção de substâncias de baixo peso molecular por gel seco de poliacrilamida e ultrafiltração sob pressão positiva. Concluiu-se que a diálise contra solução de goma-arábica, por ser preciso, relativamente exato, simples e pouco oneroso, é o mais satisfatório dentre os métodos estudados.

Four protein concentration methods, applicable to cerebrospinal fluid, were compared in order to choose a procedure that not causing protein denaturation, is accurate, precise and easy to perform in routine laboratories. The studied methods were: dialysis against arabic-gum solution, centrifuge-ultrafiltration, absorption of low molecular weight substances by dry polyacrylamide gel and pressure ultrafiltration. Blood serum was diluted, then concentrated by these procedures and analyzed by cellulose acetate electrophoresis; the undiluted serum served as control. It was observed that two methods: pressure ultrafiltration and absorption of low molecular weight substances by polyacrylamide hydrogel, are not suitable for concentrating diluted fluids because they frequently cause protein denaturation. It was concluded that dialysis against arabic-gum is the most suitable, of the methods studied, for application in routine laboratories taking in account that this procedure is precise, relatively accurate, simple and of low cost.

Proteinas do liquido cefalorraqueano: I. Estudo comparativo entre metodos de concentração

Cerebrospinal fluid proteins: I. Comparison of protein concentration methods

Lucia M. Singer Vermes

Professora Assistente do Departamento de Microbiologia e Imunologia do Instituto de Ciências Biomédicas da Universidade de São Paulo

RESUMO

No intuito de escolher entre métodos de concentração, aplicáveis ao LCR, aquele que, não provocando desnaturação de proteínas, seja o mais exato, preciso e de fácil aplicação em laboratórios de rotina, foram estudados quatro procedimentos: diálise contra solução de goma-arábica, centrífugo-ultrafiltração, absorção de substâncias de baixo peso molecular por gel seco de poliacrilamida e ultrafiltração sob pressão positiva. Concluiu-se que a diálise contra solução de goma-arábica, por ser preciso, relativamente exato, simples e pouco oneroso, é o mais satisfatório dentre os métodos estudados.

SUMMARY

Four protein concentration methods, applicable to cerebrospinal fluid, were compared in order to choose a procedure that not causing protein denaturation, is accurate, precise and easy to perform in routine laboratories. The studied methods were: dialysis against arabic-gum solution, centrifuge-ultrafiltration, absorption of low molecular weight substances by dry polyacrylamide gel and pressure ultrafiltration. Blood serum was diluted, then concentrated by these procedures and analyzed by cellulose acetate electrophoresis; the undiluted serum served as control.

It was observed that two methods: pressure ultrafiltration and absorption of low molecular weight substances by polyacrylamide hydrogel, are not suitable for concentrating diluted fluids because they frequently cause protein denaturation.

It was concluded that dialysis against arabic-gum is the most suitable, of the methods studied, for application in routine laboratories taking in account that this procedure is precise, relatively accurate, simple and of low cost.

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Full text available only in PDF format.

Resumo da tese de doutoramento "Líquido Cefalorraqueano Normal: métodos de concentração, proteinograma e imuneglobulinas", defendida no Departamento de Microbiologia e Imunologia do Instituto de Ciências Biomédicas da Universidade de São Paulo.

Departamento de Microbiologia e Imunologia - Instituto de Ciências Biomédicas - Universidade de São Paulo - Caixa Postal 2921 - 01000 - São Paulo, SP - Brasil.

1. ANGELA, G. C. - Le proteine nei liquidi biologici (siero excluso). Minerva med. 64:219, 1973.
2. COLOVER, J. - A micro technique for protein concentration suitable for quantitative electrophoresis of cerebrospinal fluid. J. clin. Path. 14:559, 1961.
3. CURTAIN, C. C. - Concentrating protein solutions. Nature, 203:1380, 1964.
4. FAULKNER, W. R.; GARDNER, M. & LEWIS, L. A. - The concentration of low-protein body fluids. Clin. Chem. 8:424, 1962.
5. GOA, J. & TVENTEN, L. - Electrophoresis of cerebrospinal fluid proteins in certain neurological disorders. Scand. J. clin. Lab. Invest 15:152, 1963.
6. GORNALL, A. G.; BARDAWILL, C. J. & DAVID, M. M. - Determination of serum proteins by means of the biuret reaction. J. biol. Chem. 177:751, 1949.
7. GREENWALT, T. J.; STEANE, E. A. & van OSS, C. J. - Improved method for concentrating urine proteins applied to the detection of Bence-Jones proteinuria. Fed. Proc. 25:612, 1966.
8. KAPLAN, A. & JOHNSTONE, M. - Concentration of cerebrospinal fluid proteins and their fractionation by cellulose acetate electrophoresis. Clin. Chem. 11:717, 1966.
9. KLEINE, T. O.; STROH, M. & STROH J. - Comparison of four differente methods to concentrate human cerebrospinal fluid. In World Congress of Anatomic and Clinical Pathology, 8ş, Munich, 1972. p. 65 (Excerpta Medica International Congress series, v. 262).
10. KOHN, J. - A new supporting medium for zone electrophoresis. Biochem. J. 65:9, 1957.
11. KOHN, J. - A simple method for the concentration of fluids containing protein. Nature 183:1055, 1959.
12. KWAPINSKY, J. B. G. - Methodology of Immunochemical and Immunological Research. Willy Interscience, New York, 1972, p. 223-224.
13. LEMMEN, L. J.; NEWMAN, N. A. & DeJONG, R. N. - Study of cerebrospinal fluid proteins with paper electrophoresis: I. a review of literature. Univ. Mich. med. Bull. 23:3, 1957.
14. LEMMEN, L. J.; TOURTELLOTTE, W. W.; GLIMN, J. G.; HIGGINS, J. E. & PARKER, J. A. - Study of cerebrospinal fluid proteins with paper electrophoresis. IV. methods for concentrating dilute protein solutions. Univ. Mich. med. Bull. 23:135, 1957.
15. MATSON, C. F. - A procedure for agar gel electrophoresis. Amer. J. clin. Pathol. 37:143, 1962.
16. McGARRY, E.; SEHON, A. H.; ROSE, B. - The isolation and electrophoretic charaterization of the proteins in the urine of normal subjects. J. clin. Invest. 34:832, 1955.
17. MEJBAUM-KATZENELLENBOGEN, W.; DOBRYSZYCKA, W.; BOGUSLAWLKA-JAWORSKA, J. & MORAWTECKA, B. - Regeneration of protein from insoluble protein-tannin compounds. Nature 184:1799, 1959.
18. SPINA-FRANÇA, A. - Eletroforese em papel das proteínas do líquido cefalorraquidiano. III. técnica. Arq. Neuro-Psiquiat. (São Paulo) 16:236, 1958.
19. VAZ, C. A. C, FERRI, R. G.; GEISHOVEL, N. & CAMPOS, A. N. P. - Eletroforese sobre acetato de celulose (CAF). Reprodutibilidade e valores normais. Rev. Inst. Adolfo Lutz (São Paulo) 31:71, 1971.
20. VERMES, L. M. S.; FERRI, R. G.; AISEN, J. & MARLET, J. M. - Proteínas totais do líquido cefalorraqueano obtido por punção da cisterna magna: valores normais (variações ligadas ao sexo). Arq. Neuro-Psiquiat. (São Paulo) 34:315, 1976.
21. WINDISCH, R. M. & BRACKEN, M, M. - Cerebrospinal fluid proteins: concentration by membrane ultrafiltration and fractionation by electrophoresis on cellulose acetate. Clin. Chem. 16:416, 1970.

Datas de Publicação

Publicação nesta coleção
14 Ago 2012
Data do Fascículo
Mar 1983

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. ANGELA, G. C. - Le proteine nei liquidi biologici (siero excluso). Minerva med. 64:219, 1973.

[2] 2. COLOVER, J. - A micro technique for protein concentration suitable for quantitative electrophoresis of cerebrospinal fluid. J. clin. Path. 14:559, 1961.

[3] 3. CURTAIN, C. C. - Concentrating protein solutions. Nature, 203:1380, 1964.

[4] 4. FAULKNER, W. R.; GARDNER, M. & LEWIS, L. A. - The concentration of low-protein body fluids. Clin. Chem. 8:424, 1962.

[5] 5. GOA, J. & TVENTEN, L. - Electrophoresis of cerebrospinal fluid proteins in certain neurological disorders. Scand. J. clin. Lab. Invest 15:152, 1963.

[6] 6. GORNALL, A. G.; BARDAWILL, C. J. & DAVID, M. M. - Determination of serum proteins by means of the biuret reaction. J. biol. Chem. 177:751, 1949.

[7] 7. GREENWALT, T. J.; STEANE, E. A. & van OSS, C. J. - Improved method for concentrating urine proteins applied to the detection of Bence-Jones proteinuria. Fed. Proc. 25:612, 1966.

[8] 8. KAPLAN, A. & JOHNSTONE, M. - Concentration of cerebrospinal fluid proteins and their fractionation by cellulose acetate electrophoresis. Clin. Chem. 11:717, 1966.

[9] 9. KLEINE, T. O.; STROH, M. & STROH J. - Comparison of four differente methods to concentrate human cerebrospinal fluid. In World Congress of Anatomic and Clinical Pathology, 8ş, Munich, 1972. p. 65 (Excerpta Medica International Congress series, v. 262).

[10] 10. KOHN, J. - A new supporting medium for zone electrophoresis. Biochem. J. 65:9, 1957.

[11] 11. KOHN, J. - A simple method for the concentration of fluids containing protein. Nature 183:1055, 1959.

[12] 12. KWAPINSKY, J. B. G. - Methodology of Immunochemical and Immunological Research. Willy Interscience, New York, 1972, p. 223-224.

[13] 13. LEMMEN, L. J.; NEWMAN, N. A. & DeJONG, R. N. - Study of cerebrospinal fluid proteins with paper electrophoresis: I. a review of literature. Univ. Mich. med. Bull. 23:3, 1957.

[14] 14. LEMMEN, L. J.; TOURTELLOTTE, W. W.; GLIMN, J. G.; HIGGINS, J. E. & PARKER, J. A. - Study of cerebrospinal fluid proteins with paper electrophoresis. IV. methods for concentrating dilute protein solutions. Univ. Mich. med. Bull. 23:135, 1957.

[15] 15. MATSON, C. F. - A procedure for agar gel electrophoresis. Amer. J. clin. Pathol. 37:143, 1962.

[16] 16. McGARRY, E.; SEHON, A. H.; ROSE, B. - The isolation and electrophoretic charaterization of the proteins in the urine of normal subjects. J. clin. Invest. 34:832, 1955.

[17] 17. MEJBAUM-KATZENELLENBOGEN, W.; DOBRYSZYCKA, W.; BOGUSLAWLKA-JAWORSKA, J. & MORAWTECKA, B. - Regeneration of protein from insoluble protein-tannin compounds. Nature 184:1799, 1959.

[18] 18. SPINA-FRANÇA, A. - Eletroforese em papel das proteínas do líquido cefalorraquidiano. III. técnica. Arq. Neuro-Psiquiat. (São Paulo) 16:236, 1958.

[19] 19. VAZ, C. A. C, FERRI, R. G.; GEISHOVEL, N. & CAMPOS, A. N. P. - Eletroforese sobre acetato de celulose (CAF). Reprodutibilidade e valores normais. Rev. Inst. Adolfo Lutz (São Paulo) 31:71, 1971.

[20] 20. VERMES, L. M. S.; FERRI, R. G.; AISEN, J. & MARLET, J. M. - Proteínas totais do líquido cefalorraqueano obtido por punção da cisterna magna: valores normais (variações ligadas ao sexo). Arq. Neuro-Psiquiat. (São Paulo) 34:315, 1976.

[21] 21. WINDISCH, R. M. & BRACKEN, M, M. - Cerebrospinal fluid proteins: concentration by membrane ultrafiltration and fractionation by electrophoresis on cellulose acetate. Clin. Chem. 16:416, 1970.