Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi

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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 wit...

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Detalles Bibliográficos
Autor principal: Kaplan, D.
Otros Autores: Baldi, C., Chiaramonte, M.G, Fernandez, M.M, Levin, M.J, Malchiodi, E. L., Baldi, A.
Formato: Capítulo de libro
Lenguaje:Inglés
Publicado: Academic Press Inc. 1998
Acceso en línea:Registro en Scopus
DOI
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Aporte de:Registro referencial: Solicitar el recurso aquí
Biblioteca Central Dr. Luis F. Leloir (FCEN) de Universidad de Buenos Aires
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024 7 |2 Molecular Sequence Numbers  |a GENBANK: AF091431; 
024 7 |2 cas  |a cruzipain; cysteine proteinase, 37353-41-6; RNA, 63231-63-0 
030 |a BBRCA 
040 |a Scopus  |b spa  |c AR-BaUEN  |d AR-BaUEN 
100 1 |a Kaplan, D. 
245 1 0 |a Expression of a recombinant Fab antibody fragment against cruzipain, the major cysteine proteinase of Trypanosoma cruzi 
260 |b Academic Press Inc.  |c 1998 
270 1 0 |m Baldi, A.; IBYME, Obligado 2490, (1428) Buenos Aires, Argentina; email: abaldi@proteus.dna.uba.ar 
504 |a Brener, Z., (1982) Bull. World Health Organ., 60, pp. 463-473 
504 |a (1984) World Health Organ. Tech. Report. Ser., 701, pp. 53-60 
504 |a Chiller, T.M., Samudio, M.A., Zoulek, G., (1990) Am. J. Trop. Med. Hyg., 22, pp. 696-698 
504 |a Malchiodi, E.L., Chiaramonte, M.G., Taranto, N.J., Zwirner, N.W., Margni, R.A., (1994) Clin. Exp. Immunol., 97, pp. 417-423 
504 |a Camargo, M.E., (1988) Laboratory Diagnosis of Infectious Disease, Principles and Practice, 1, pp. 744-753. , Springer Verlag, New York 
504 |a Schattschneider, W., Lopes, E.R., De Alencar, J.E., (1992) Trop. Geogr. Med., 44, pp. 210-218 
504 |a 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 
504 |a 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 
504 |a Martinez, J., Campetella, O., Frasch, A.C.C., Cazzulo, J.J., (1991) Infect. Immunity, 59, p. 4275 
504 |a 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 
504 |a 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 
504 |a Barbas C.F. III, Kang, A.S., Lerner, R.A., Benkovic, S.J., (1991) Proc. Natl. Acad. Sci. USA, 88, pp. 7978-7982 
504 |a 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 
504 |a 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 
504 |a Goldbaum, F.A., Leoni, J., Wallace, J.C., Fossati, C.A., (1993) J. Clin. Microbiol., 31, pp. 2141-2145 
504 |a 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 
504 |a 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 
504 |a Viotti, R., Vigliano, C., Armenti, H., Segura, E., (1994) Am. Heart J., 127, pp. 151-162 
504 |a Franke De Cazzulo, B.M., Martinez, J., North, M.J., Coombs, G.H., Cazzulo, J.J., (1994) FEMS Microbiol. Lett., 124, pp. 81-86 
504 |a 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 
504 |a Cazzulo, J.J., Stoka, V., Turk, V., (1997) J. Biol. Chem., 378, pp. 1-10 
504 |a Gonzalez, G., Orn, A., Cazzulo, J.J., Gronvik, K.O., (1994) Scand. J. Immunol., 40, pp. 389-394 
504 |a 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 
504 |a Lalmanach, G., Mayer, R., Serveau, C., Scharfstein, J., Gauthier, F., (1996) Biochem. J., 318, pp. 395-399 
506 |2 openaire  |e Política editorial 
520 3 |a 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.  |l eng 
536 |a Detalles de la financiación: Universidad de Buenos Aires, UBA 
536 |a Detalles de la financiación: National Research Council, NRC 
536 |a Detalles de la financiación: This work was financially supported by National Research Council (CONICET-Argentina) and University of Buenos Aires (UBA). 
593 |a Inst. of Biol./Exp. Medicine (IBYME), Obligado 2490, 1428, Buenos Aires, Argentina 
593 |a Inst. of Humoral Immunity Studies, IDEHU, CONICET-UBA, Buenos Aires, Argentina 
593 |a Inst. of Genetic Eng./Molec. Biology, CONICET-UBA, Buenos Aires, Argentina 
690 1 0 |a COMPLEMENTARY DNA 
690 1 0 |a CRUZIPAIN 
690 1 0 |a CYSTEINE PROTEINASE 
690 1 0 |a ENZYME ANTIBODY 
690 1 0 |a IMMUNOGLOBULIN F(AB) FRAGMENT 
690 1 0 |a IMMUNOGLOBULIN HEAVY CHAIN 
690 1 0 |a IMMUNOGLOBULIN KAPPA CHAIN 
690 1 0 |a PARASITE ANTIBODY 
690 1 0 |a PARASITE ANTIGEN 
690 1 0 |a PRIMER DNA 
690 1 0 |a RECOMBINANT ANTIBODY 
690 1 0 |a RNA 
690 1 0 |a ANIMAL CELL 
690 1 0 |a ANTIGEN RECOGNITION 
690 1 0 |a ARTICLE 
690 1 0 |a CONTROLLED STUDY 
690 1 0 |a ENZYME LINKED IMMUNOSORBENT ASSAY 
690 1 0 |a ESCHERICHIA COLI 
690 1 0 |a GENE EXPRESSION SYSTEM 
690 1 0 |a HYBRIDOMA 
690 1 0 |a IMMUNOFLUORESCENCE 
690 1 0 |a MOLECULAR CLONING 
690 1 0 |a NONHUMAN 
690 1 0 |a NUCLEOTIDE SEQUENCE 
690 1 0 |a PRIORITY JOURNAL 
690 1 0 |a TRYPANOSOMA CRUZI 
690 1 0 |a ANIMALIA 
690 1 0 |a ESCHERICHIA COLI 
690 1 0 |a ESCHERICHIA COLI 
690 1 0 |a TRYPANOSOMA 
690 1 0 |a TRYPANOSOMA CRUZI 
690 1 0 |a TRYPANOSOMA CRUZI 
700 1 |a Baldi, C. 
700 1 |a Chiaramonte, M.G. 
700 1 |a Fernandez, M.M. 
700 1 |a Levin, M.J. 
700 1 |a Malchiodi, E. L. 
700 1 |a Baldi, A. 
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