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Abstract:

In the present study we describe the production and characterization of a panel of monoclonal antibodies (MoAbs) directed against cruzipain (Crz), the major cysteine proteinase from Trypanosoma cruzi. The five MoAbs, BD6, BF2, CG2, CH8, and DC10 were analysed with respect to affinity and specificity. None of the MoAbs cross‐reacted with papain, which has regions of high homology with Crz. Treatment of the antigen with periodate did not affect the binding of the MoAbs. suggesting that they bind to the polypeptide moiety of Crz. CH8 recognized a continuous epitope located at the C‐terminal extension of the proteinase that appeared to be highly immunogenic. Although the rest of the MoAbs recognized epitopes located in the catalytic domain, the enzymatic activity of Crz was not impaired by the binding of the MoAbs. Characterization of the antibody‐binding sites revealed the presence of at least four separate epitopes. Copyright © 1994, Wiley Blackwell. All rights reserved

Registro:

Documento: Artículo
Título:Production and Characterization of Monoclonal Antibodies Against the Major Cysteine Proteinase of Trypanosoma cruzi
Autor:GONZALEZ, G.; ÖRN, A.; CAZZULO, J.J.; GRÖNVIK, K.‐O.
Filiación:Cátedra de Inmunologia, Facultad de Quimica, Universidad de la República, Montevideo, Uruguay
Department of Immunology, Karolinska Institute, Stockholm, Sweden
Fundación Campomar, Facultad de Ciencias Exactas Y Naturales, UBA, Buenos Aires, Argentina
Section of Immunology A Cell Culture, National Veterinary Institute, BMC, Uppsala, Sweden
Palabras clave:cruzipain; cysteine proteinase; monoclonal antibody; parasite antigen; animal experiment; article; enzyme activity; enzyme linked immunosorbent assay; mouse; nonhuman; trypanosoma cruzi; Animal; Antibodies, Monoclonal; Antibodies, Protozoan; Antibody Affinity; Antibody Specificity; Cysteine Endopeptidases; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Mice; Mice, Inbred BALB C; Precipitin Tests; Support, Non-U.S. Gov't; Trypanosoma cruzi
Año:1994
Volumen:40
Número:4
Página de inicio:389
Página de fin:394
DOI: http://dx.doi.org/10.1111/j.1365-3083.1994.tb03479.x
Título revista:Scandinavian Journal of Immunology
Título revista abreviado:Scand. J. Immunol.
ISSN:03009475
CAS:Antibodies, Monoclonal; Antibodies, Protozoan; cruzipain, EC 3.4.22.-; Cysteine Endopeptidases, EC 3.4.22
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03009475_v40_n4_p389_GONZALEZ

Referencias:

  • Cazzulo, JJ, Frasch, AC, SAPA/trans‐sialidase and cruzipain: two antigens from Trypanosoma cruzi contain immunodominant but enzymatically inactive domains (1992) FASEB J, 6, pp. 3259-3264
  • Souto‐Padron, T, Campetella, O, Cazzulo, JJ, De Souza, W, Cysteine proteinases in Trypanosoma cruzi immuno‐cytochemical localization and involvement in parasite‐host cell interaction (1990) Cell Sci, 96, pp. 485-490
  • Åslund, L, Henriksson, J, Campetella, O, Frasch, AC, Pettersson, U, Cazzulo, JJ, The C‐terminal extension of the major cysteine proteinase (cruzipain) from Trypanosoma cruzi (1991) Molecular and Biochemical Parasitology, 45, pp. 345-348
  • Martinez, J, Campetella, O, Frasch, AC, Cazzulo, JJ, The reactivity of sera from chagasic patients against different fragments of cruzipain, the major cysteine proteinase from Trypanosoma cruzi, suggests the presence of defined antigenic and catalytic domains (1993) Immunol Lett, 35, pp. 191-196
  • Cazzulo, JJ, Couso, R, Raimondi, A, Wemstedt, C, Hellman, U, Further characterization and partial amino acid sequence of a cysteine proteinase from Trypanosoma cruzi (1989) Mol Biochem Parasitol, 33, pp. 33-42
  • Laemmli, UK, Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685
  • Tijssen, P, Preparation of enzyme‐antibody or other enzyme‐macromolecule conjugates (1985) Practice and Theory of Enzyme lmmunoassays, pp. 221-278. , Burdon RH, Knippenberg PH,. New York:, Elsevier Science Publishers
  • van Heyningen, V, Brock, DJ, van Heyningen, S, A simple method for ranking the affinity of monoclonal antibodies (1983) J Immunol Methods, 62, pp. 147-153
  • Cazzulo, JJ, Martinez, J, Parodi, AJ, Wemstedt, C, Hellman, U, On the post‐transiational modifications at the C‐terminal domain of the major cysteine proteinase (cruzipain) from Trypanosoma cruzi (1992) FEMS Microbiol Lett, 100, pp. 411-416
  • Martinez, J, Campetells, O, Frasch, AC, Cazzulo, JJ, The major cysteine proteinase (cruzipain) from trypanosoma cruzi is antigenic in human infections (1991) Infect Immun, 59, pp. 4275-4277
  • Cazzulo, JJ, Hellman, U, Couso, R, Paradodi, AJ, Amino acid and carbohydrate composition of a Lysosomal cysteine proteinase from Trypanosoma Cruzi. Absence of phosphorylated man nose residues (1990) Mol Biochem Parasitol, 38, pp. 41-48

Citas:

---------- APA ----------
GONZALEZ, G., ÖRN, A., CAZZULO, J.J. & GRÖNVIK, K.‐O. (1994) . Production and Characterization of Monoclonal Antibodies Against the Major Cysteine Proteinase of Trypanosoma cruzi. Scandinavian Journal of Immunology, 40(4), 389-394.
http://dx.doi.org/10.1111/j.1365-3083.1994.tb03479.x
---------- CHICAGO ----------
GONZALEZ, G., ÖRN, A., CAZZULO, J.J., GRÖNVIK, K.‐O. "Production and Characterization of Monoclonal Antibodies Against the Major Cysteine Proteinase of Trypanosoma cruzi" . Scandinavian Journal of Immunology 40, no. 4 (1994) : 389-394.
http://dx.doi.org/10.1111/j.1365-3083.1994.tb03479.x
---------- MLA ----------
GONZALEZ, G., ÖRN, A., CAZZULO, J.J., GRÖNVIK, K.‐O. "Production and Characterization of Monoclonal Antibodies Against the Major Cysteine Proteinase of Trypanosoma cruzi" . Scandinavian Journal of Immunology, vol. 40, no. 4, 1994, pp. 389-394.
http://dx.doi.org/10.1111/j.1365-3083.1994.tb03479.x
---------- VANCOUVER ----------
GONZALEZ, G., ÖRN, A., CAZZULO, J.J., GRÖNVIK, K.‐O. Production and Characterization of Monoclonal Antibodies Against the Major Cysteine Proteinase of Trypanosoma cruzi. Scand. J. Immunol. 1994;40(4):389-394.
http://dx.doi.org/10.1111/j.1365-3083.1994.tb03479.x