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Kaplan, D.; Baldi, C.; Chiaramonte, M.G.; Fernandez, M.M.; Levin, M.J.; Malchiodi, E.; Baldi, A. "Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi" (1998) Biochemical and Biophysical Research Communications. 253(1):53-58
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Abstract:

Cruzipain, the major proteinase of Trypanosoma cruzi, plays an important role in the biology of this parasite. This study reports the development of a recombinant Fab antibody, using RNA isolated from the anti-Ag163B6 hybridoma against cruzipain. This procedure involves the use of cDNAs obtained with the aid of a specific set of primers complementary to the complete light kappa chain (Lκ) and the first two domains of the IgG1 heavy chain (VH/CH1). These products were subsequently cloned in the pComb3 system, from which the gIII gene had been removed, and expressed in Escherichia coli cells. The recombinant Fab molecule recognized cruzipain by ELISA, in a fashion similar to the original mAb anti Ag163B6. Nucleotide sequence analysis of the recombinant molecule, together with its immunological recognition by specific anti-mouse IgG (Fab)2, indicated the immunoglobulin nature of the recombinant product. Moreover, both the mAb anti-Ag163B6 and the soluble Fab fragment described here react similarly with the intact parasite surface, as observed in an indirect immunofluorescence assay. In conclusion, our recombinant Fab anti-Ag163B6 allows the possible use of this molecule for diagnosis, antigen purification, and eventually treatment of Chagas-afflicted individuals.

Registro:

Documento: Artículo
Título:Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi
Autor:Kaplan, D.; Baldi, C.; Chiaramonte, M.G.; Fernandez, M.M.; Levin, M.J.; Malchiodi, E.; Baldi, A.
Filiación:Inst. of Biol./Exp. Medicine (IBYME), Obligado 2490, 1428, Buenos Aires, Argentina
Inst. of Humoral Immunity Studies, IDEHU, CONICET-UBA, Buenos Aires, Argentina
Inst. of Genetic Eng./Molec. Biology, CONICET-UBA, Buenos Aires, Argentina
Palabras clave:complementary DNA; cruzipain; cysteine proteinase; enzyme antibody; immunoglobulin F(ab) fragment; immunoglobulin heavy chain; immunoglobulin kappa chain; parasite antibody; parasite antigen; primer DNA; recombinant antibody; RNA; animal cell; antigen recognition; article; controlled study; enzyme linked immunosorbent assay; Escherichia coli; gene expression system; hybridoma; immunofluorescence; molecular cloning; nonhuman; nucleotide sequence; priority journal; Trypanosoma cruzi; Animalia; Escherichia coli; Escherichia coli; Trypanosoma; Trypanosoma cruzi; Trypanosoma cruzi
Año:1998
Volumen:253
Número:1
Página de inicio:53
Página de fin:58
DOI: http://dx.doi.org/10.1006/bbrc.1998.9654
Título revista:Biochemical and Biophysical Research Communications
Título revista abreviado:Biochem. Biophys. Res. Commun.
ISSN:0006291X
CODEN:BBRCA
CAS:cruzipain; cysteine proteinase, 37353-41-6; RNA, 63231-63-0
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0006291X_v253_n1_p53_Kaplan

Referencias:

  • Brener, Z., (1982) Bull. World Health Organ., 60, pp. 463-473
  • (1984) World Health Organ. Tech. Report. Ser., 701, pp. 53-60
  • Chiller, T.M., Samudio, M.A., Zoulek, G., (1990) Am. J. Trop. Med. Hyg., 22, pp. 696-698
  • Malchiodi, E.L., Chiaramonte, M.G., Taranto, N.J., Zwirner, N.W., Margni, R.A., (1994) Clin. Exp. Immunol., 97, pp. 417-423
  • Camargo, M.E., (1988) Laboratory Diagnosis of Infectious Disease, Principles and Practice, 1, pp. 744-753. , Springer Verlag, New York
  • Schattschneider, W., Lopes, E.R., De Alencar, J.E., (1992) Trop. Geogr. Med., 44, pp. 210-218
  • Chiaramonte, M.G., Zwirner, N.W., Caropresi, S.L., Taranto, N.J., Malchiodi, E.L., (1996) Am. J. Trop. Med. Hyg., 54 (3), pp. 271-273
  • Malchiodi, E.L., Chiaramonte, M.G., Martinez, J.A., Zwirner, N.W., Margni, R.A., Cazzulo, J.J., (1993) Immunol. Lett., 35, pp. 59-62
  • Martinez, J., Campetella, O., Frasch, A.C.C., Cazzulo, J.J., (1991) Infect. Immunity, 59, p. 4275
  • Carbonetto, C.H., Malchiodi, E.L., Chiaramonte, M.G., Durante De Isola, E., Fossati, C.A., Margni, R.A., (1990) Clin. Exp. Immunol., 82, pp. 93-96
  • Cabilly, S., Rigss, A.D., Pande, H., Shively, J.E., Holmes, W.E., Rey, M., Perry, L.J., Heyneker, H.L., (1984) Proc. Natl. Acad. Sci. USA, 81, pp. 3273-3277
  • Barbas C.F. III, Kang, A.S., Lerner, R.A., Benkovic, S.J., (1991) Proc. Natl. Acad. Sci. USA, 88, pp. 7978-7982
  • Huse, W.D., Sastry, L., Iverson, S.A., Kang, A.S., Alting-Mees, M., Burton, D.R., Benkovic, S.J., Lerner, R.A., (1989) Science, 246, pp. 1275-1281
  • Sambrook, J., Fritsch, E.F., Maniatis, T., (1989), Cold Spring Harbor Laboratory Press; Alvarez, M., Cerisola, J.A., Rohwedder, R.W., (1968) Bol. Chile Parasitol., 23, pp. 4-9
  • Goldbaum, F.A., Leoni, J., Wallace, J.C., Fossati, C.A., (1993) J. Clin. Microbiol., 31, pp. 2141-2145
  • Levin, M.J., Franco Da Silveira, J., Frasch, A.C.C., Camargo, M.E., Lafon, S., Degrave, W.M., Rangel Aldao, R., (1991) FEMS Microbiol. Immunol., 89, pp. 11-20
  • Kaplan, D., Ferrari, I., Lopez Bergami, P., Mahler, E., Levitus, G., Chiale, P., Hoebeke, J., Levin, M., (1997) Proc. Natl. Acad. Sci. USA, 94, pp. 10301-10306
  • Viotti, R., Vigliano, C., Armenti, H., Segura, E., (1994) Am. Heart J., 127, pp. 151-162
  • Franke De Cazzulo, B.M., Martinez, J., North, M.J., Coombs, G.H., Cazzulo, J.J., (1994) FEMS Microbiol. Lett., 124, pp. 81-86
  • Meirelles, M.N., Juliano, L., Carmona, E., Silva, S.G., Costa, E.M., Murta, A.C., Scharfstein, J., (1992) Mol. Biochem. Parasitol., 52, pp. 175-184
  • Cazzulo, J.J., Stoka, V., Turk, V., (1997) J. Biol. Chem., 378, pp. 1-10
  • Gonzalez, G., Orn, A., Cazzulo, J.J., Gronvik, K.O., (1994) Scand. J. Immunol., 40, pp. 389-394
  • McGrath, M.E., Eakin, A.E., Engel, J.C., McKerrow, J.H., Craik, C.S., Fletterick, R.J., (1995) J. Mol. Biol., 247, pp. 251-259
  • Lalmanach, G., Mayer, R., Serveau, C., Scharfstein, J., Gauthier, F., (1996) Biochem. J., 318, pp. 395-399

Citas:

---------- APA ----------
Kaplan, D., Baldi, C., Chiaramonte, M.G., Fernandez, M.M., Levin, M.J., Malchiodi, E. & Baldi, A. (1998) . Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi. Biochemical and Biophysical Research Communications, 253(1), 53-58.
http://dx.doi.org/10.1006/bbrc.1998.9654
---------- CHICAGO ----------
Kaplan, D., Baldi, C., Chiaramonte, M.G., Fernandez, M.M., Levin, M.J., Malchiodi, E., et al. "Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi" . Biochemical and Biophysical Research Communications 253, no. 1 (1998) : 53-58.
http://dx.doi.org/10.1006/bbrc.1998.9654
---------- MLA ----------
Kaplan, D., Baldi, C., Chiaramonte, M.G., Fernandez, M.M., Levin, M.J., Malchiodi, E., et al. "Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi" . Biochemical and Biophysical Research Communications, vol. 253, no. 1, 1998, pp. 53-58.
http://dx.doi.org/10.1006/bbrc.1998.9654
---------- VANCOUVER ----------
Kaplan, D., Baldi, C., Chiaramonte, M.G., Fernandez, M.M., Levin, M.J., Malchiodi, E., et al. Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi. Biochem. Biophys. Res. Commun. 1998;253(1):53-58.
http://dx.doi.org/10.1006/bbrc.1998.9654