Summary
The extraction and purification of gangliosides from brains of animals infected with the scrapie agent was evaluated by scaling-down a large-scale procedure currently used with bovine brains. Inactivation experiments employed hamster brains infected with the 263 K strain of scrapie. Residual infectivity was determined at different points of the procedure and in the final preparation by an in vivo animal bioassay. The efficacy of single steps, which included chemicals or physical techniques known to affect the viability of the scrapie agent, was verified by spiking experiments where known amounts of infectivity were added just before each single step. Infectivity was significantly diminished at early stages of the purification, and no infectivity was detected in the final preparation containing purified gangliosides. The significant reduction of infectivity at intermediate steps and the total absence of detectable infectivity in the final product confirmed that a combination of heat and exposure to sodium hydroxide, even in the presence of organic solvents, completely inactivated the scrapie agent.
Similar content being viewed by others
References
Asher DM, Gibbs CJ Jr, Gajdusek DC (1986) Slow viral infections: safe handling of the agents of subacute spongiform encephalopathies. In: Miller BM (ed) Laboratory safety: principles and practices. American Society for Microbiology, Washington, DC, pp 59–71
Brown P (1985) Virus sterility for human growth hormone. Lancet ii: 729–730
Brown P, Cathala F, Raubertas RF, Gajdusek DC, Castaigne P (1987) The epidemiology of Creutzfeldt-Jakob disease: conclusion of a 15-year investigation in France and review of the world literature. Neurology 37: 895–904
Brown P, Gajdusek DC, Gibbs CJ Jr, Asher DM (1985) Potential epidemic of Creutzfeldt-Jakob disease from human growth hormone therapy. N Engl J Med 313: 728–731
Brown P, Rohwer RG, Gajdusek DC (1984) Sodium hydroxide decontamination of Creutzfeldt-Jakob disease virus. N Engl J Med 310: 727
Brown P, Rohwer RG, Gajdusek DC (1986) Newer data on the inactivation of scrapie virus or Creutzfeldt-Jakob disease virus in brain tissue. J Infect Dis 153: 1145–1148
Chigorno V, Sonnino S, Ghidoni R, Tettamanti G (1982) Densitometric quantification of brain gangliosides separated by two-dimensional thin-layer chromatography. Neurochem Int 5: 397–403
Formisano S, Johnson ML, Lee G, Aloj SM, Edelhock H (1979) Critical micelle concentrations of gangliosides. Biochemistry 18: 1119–1124
Gajdusek DC (1990) Subacute spongiform encephalopathies: transmissible cerebral amyloidoses caused by unconventional viruses. In: BN Fields, DM Knipe (eds) Virology, 2nd edn. Raven Press, New York, pp 2289–2324
Geisler FH, Dorsey FC, Coleman WP (1991) Motor recovery following human spinal cord injury. A randomized placebo controlled trial with GM1 ganglioside. N Engl J Med 324: 1829–1838
Hope J, Reekie LJD, Hunter N, Multhaup G, Beyreuther K, White H, Scott AC, Stack MJ, Dawson M, Wells GAH (1988) Fibrils from brain of cows with new cattle disease contain scrapie-associated protein. Nature 336: 390–392
IUPAC-IUB Commission on biochemical nomenclature (1977) The nomenclature of lipids. Eur J Biochem 79: 11–21
Kimberlin RH, Walker CA (1989) Transport, targeting and replication of scrapie in CNS. In: Court LA, Dormont D, Brown P, Kingsbury DT (eds) Unconventional virus diseases of the central nervous system. Commissariat à l'Énergie Atomique, Fontenayaux-Roses, France, pp 547–562
Kimberlin RH, Walker CA, Millson GC, Taylor DM, Robertson PA, Tomlinson AH, Dickinson AG (1983) Disinfection studies with two strain of mouse-passaged scrapie agent. J Neurol Sci 59: 355–369
Masters CL, Harris JO, Gajdusek DC, Gibbs CJ Jr, Bernouilli C, Asher DM (1979) Creutzfeldt-Jakob disease: patterns of world-wide occurrence and the significance of familial and sporadic clustering. Ann Neurol 5: 177–188
Minor P (1990) Bovine spongiform encephalopathy and biological products for human use. Biologicals 18: 77–80
Morgan KL (1988) Bovine spongiform encephalopathy: time to take scrapie seriously. Vet Rec 122: 445–446
Parry HB (1983) Scrapie disease in sheep. Academic Press, London
Pocchiari M, Peano S, Conz A, Eshkol A, Maillard F, Brown P, Gibbs CJ Jr, Xi YG, Tenham-Fisher E, Macchi G (1991) Combination ultrafiltration and 6M urea treatment of human growth hormone minimizes risk from potential CJD contamination. Horm Res 35: 161–166
Prusiner SB (1991) Molecular biology of prion diseases. Science 252: 1515–1522
Prusiner SB, Groth DF, McKinley MP, Cochran SP, Bowman KA, Casper KC (1981) Thiocyanate and hydroxyl ions inactivate the scrapie agent. Proc Natl Acad Sci USA 78: 4606–4610
Reed LJ, Muench H (1938) A simple method of estimating fifty per cent endpoints. Am J Hyg 27: 493–497
Rosenberg RN, White CL, Brown P, Gajdusek DC, Volpe JJ, Posner J, Dyck PJ (1986) Precautions in handling tissues, fluids, and other contaminated material from patients with documented or suspected Creuzfeldt-Jakob disease. Ann Neurol 19: 75–77
Sabel BA, Stein DG (1986) Pharmacological treatment of central nervous system injury. Nature 323: 493
Safar J, Ceroni M, Piccardo P, Gajdusek DC, Gibbs CJ Jr (1990) Scrapie-associated precursor proteins: antigenic relationship between species and immunochemical localization in normal, scrapie and Creutzfeldt-Jakob disease brains. Neurology 40: 513–517
Safar J, Ceroni M, Piccardo P, Liberski PP, Miyazaki M, Gajdusek DC, Gibbs CJ Jr (1990) Subcellular distribution and physicochemical properties of scrapie-associated precursor protein and relationship with the scrapie agent. Neurology 40: 503–508
Svennerholm L, Fredman P (1980) A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta 617: 97–109
Tamai Y, Taguchi F, Miura S (1988) Inactivation of the Creutzfeldt-Jakob disease agent. Ann Neurol 24: 466–467
Tateishi J, Tashima T, Kitamoto T (1988) Inactivation of the Creutzfeldt-Jakob disease agent. Ann Neurol 24: 466
Walker AS, Inderlied CB, Kingsbury DT (1983) Conditions for the chemical and physical inactivation of the K.Fu. strain of the agent of Creutzfeldt-Jakob disease. Am J Public Health 73: 661–665
Wells GAH, Scott AC, Johnson CT, Gunning RF, Hancock RD, Jeffrey M, Dawson M, Bradley R (1987) A novel progressive spongiform encephalopathy in cattle. Vet Rec 121: 419–420
Wiegandt H (1985) Gangliosides. In: Wiegandt H (ed) Glycolipids. Elsevier, Amsterdam, pp 199–260 (New comprehensive biochemistry, vol 10)
Wilesmith JW (1990) Epidemiology and current status of bovine spongiform encephalopathy in the United Kingdom. J Am Vet Med Assoc 196: 1674–1675
Yu RK, Saito M (1989) Structure and localization of gangliosides. In: Margolis RU, Margolis RK (eds) Neurobiology of glycoconjugates. Plenum, New York, pp 1–42
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Di Martino, A., Safar, J., Ceroni, M. et al. Purification of non-infectious ganglioside preparations from scrapie-infected brain tissue. Archives of Virology 124, 111–121 (1992). https://doi.org/10.1007/BF01314629
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01314629