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2016, Number 3

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VacciMonitor 2016; 25 (3)

Increase in processing capacity for N-Glicolil GM3 production

Gutiérrez-Anillo JM, González-Lavaut JA, González-Villanueva L, Navarrete-Pita Y, Cancio-Méndez J, García-González N

Language: Spanish
References: 17
Page: 60-67
PDF size: 345.46 Kb.


ABSTRACT

The N-Glicolil GM3 is a monosialilganglioside used as antigen and adjuvant in vaccines preparations for therapeutic use against breast, lung and melanoma cancer. The obtaining process of this ganglioside from equine erythrocytes comprises two stages: extraction and purifi cation, in which we work to increment processing volumes. The assessment of increment in the processing capacity, its influence on the performance and the evaluation of purification process behavior by Preparative High-Performance Liquid Chromatography (Prep HPLC) constituted the objectives of this paper. The results obtained from the approved procedure and the proposal of change were compared. The process was controlled by evaluating the organoleptic characteristics and weight of lyophilized extract, as well as the detection and content of N-Glicolyl GM3 by Qualitative Thin Layer Chromatography and colorimetric analysis of sialic acid; STATGRAPHICS software was used to assess the process variability. The increment in processing volume regarding the established procedure remained the extraction stage performance and incremented slightly the purification performance, by reducing the processing time with the use of Prep HPLC. The increment of processing capacity maintained the overall performance of the process in 0,006 g of N-Glicolyl GM3/g of erythrocytes and the product obtained by means of the proposed method fulfilled the quality specifications established for the same; allowing an increment in productivity by 30%, less energy consumption and less exhaustion of human resources, being feasible to implement the change.


REFERENCES

  1. Mulens V, Marinello P, Carr A, Mazorra Z, Fernández LE. Gangliósidos en la Biología e Inmunoterapia del Cáncer: la Experiencia Cubana. Cancerología 2009;4:155-67.

  2. Zancada L. Estudio del contenido de oligosacáridos, glicolípidos y fosfolípidos de la leche de oveja. Participación en la defensa del recién nacido frente a infecciones [Tesis Doctoral]. Salamanca: Departamento de Bioquímica y Biología Molecular, Universidad de Salamanca; 2008.

  3. Bada A, Casacó A, Arteaga M, Martínez J, León A, Santana E, et al. Toxicity of a GM3 cancer vaccine in Macaca fascicularis monkey: a 12-month study. Human & Experimental Toxicology 2002;21(5):263-7.

  4. Hanais N, Dohi T, Norese G, Hakomori S. A Novel Ganglioside, De-N-acetyl-GM3 (II3NeuNH2LacCer), acting as a Strong Promoter for Epidermal Growth Factor Receptor Kinase and as a Stimulator for Cell Growth. The Journal of Biological Chemistry 1988;263(13):6296-301.

  5. Izquierdo N, Lorizate M, Contreras FX, Rodríguez MT, Glass B, Erkizia I, et al. Sialyllactose in Viral Membrane Gangliosides Is a Novel Molecular Recognition Pattern for Mature Dendritic Cell Capture of HIV-1. PLoS Biol 2012;10(4):e1001315. doi:10.1371/ journal.pbio.1001315

  6. Puente R. Gangliósidos de la leche: variaciones de su contenido en diferentes etapas de la lactación. Estudio de su posible función. [Tesis Doctoral]. León: Facultad de Biología, Universidad de León; 1992.

  7. Regalado A, Mesa M, Chávez D, González R, Monteserín LA, Rodríguez MC, et al. A New and Effi cient Approach to Prepare N-acetyl GM Ganglioside Via Trisaccharide [1→4] Lactone. Organic Process Research & Development 2013;17(1):53–60.

  8. Mazorra Z. El gangliósido GM3 como blanco en la inmunoterapia del melanoma. [Tesis Doctoral]. La Habana: Centro de Inmunología Molecular; 2007.

  9. Garrido R, Veloso RC, López M, González JA, Rodríguez MC, Vélez H, et al. Evaluation and evidences of natural gangliosides with two unsaturated bonds in the ceramide structure by combination of MALDI-MS and NMR. Anal Bioanal. Chem. 2011;400(10):3675-80.

  10. Carr A, Mazorra Z, Alonso DF, Mesa C, Valiente O, Gómez DE, et al. A purifi ed GM3 ganglioside conjugated vaccine induces specifi c, adjuvant-dependent and non-transient antitumour activity against B16 mouse melanoma in vitro and in vivo. Melanoma Research 2001;11:219-27.

  11. Arango MC, González A. Vacunas terapéuticas en cáncer: Ensayos clínicos actuales. Revista Cubana de Medicina 2002;41(6). Disponible en: http://scielo.sld.cu/scielo.php?script=sci_ arttext&pid=S0034-75232002000600009&lng=es.

  12. Portoukalian J, Carrel S, Doré JF, Rümke P. Humoral Immune Response in Disease-free Advanced Melanoma Patients after Vaccination with Melanoma- associated Gangliosides. Int. J. Cancer 1991;49(6):893-9.

  13. Alvarez N, Desselle A, Cochonneau D, Chaumette T, Clemenceau B, et al. A Monoclonal Antibody to O-Acetyl-GD2 Ganglioside and Not to GD2 Shows Potent Anti-Tumor Activity without Peripheral Nervous System Cross-Reactivity. PLoS ONE 2011;6(9):e25220. doi: 10.1371/journal.pone.0025220

  14. Folch-Pi J, Arsove S, Meath JA. Isolation of Brain Strandin, a New Type of Large Molecular Tissue Component. The Journal of Biological Chemistry 1951;191:819-31.

  15. Wang WQ, Gustafson A. Erythrocyte Lipid Extraction in Alcohol- Chloroform Ganglioside Systems: A Comparative Study. Acta Chemica Scandinavica 1994;48:753-8.

  16. Wang WQ, Gustafson A. Ganglioside Extraction from Erythrocytes: a Comparison Study. Acta Chemica Scandinavica 1995;49:929-36.

  17. Svennerholm L. Quantitative estimation of sialic acids. A colorimetric resorcinol-hydrochloric acid method. Biochimica et Biophysica Acta 1957;24:604-11.




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