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Specific antibody immune response against the parasitic portion of a glutathione‐S‐transferase fusion protein
Author(s) -
LópezMonteon Aracely,
RamosLigonio Angel,
PérezCastillo Leticia,
TalamásRohana Patricia,
Luis RosalesEncina José
Publication year - 2003
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.02-0471com
Subject(s) - antibody , fusion protein , biology , microbiology and biotechnology , immune system , recombinant dna , antibody titer , titer , glutathione s transferase , immunology , glutathione , gene , biochemistry , enzyme
The humoral immune response against an Entamoeba histolytica recombinant protein has been investigated. The 628 bp Bam HI ‐Eco RI DNA fragment (L1b) from the M11 cDNA clone, partially coding for a 220 kDa (L220) protein, was ligated in‐frame into the pGEX‐3X plasmid vector to produce the fusion protein GST‐L1b. BALB/c mice were immunized with different doses of the GST‐L1b fusion protein (10–500 µg). GST‐L1b doses of 100/50/50, 300, or 500 µg induced an antibody response (IgG1>IgG3, IgG2a>IgG2b) specific for the amoebic part of the fusion protein (L1b). These antibodies were able to recognize the native protein in amoebic total extract. Anti‐GST antibodies were not detected. On the other hand, doses of 10/ 10/10 or 200/100/100 µg induced antibodies able to recognize both GST (IgG2a>IgG1>IgG2b) and L1b (IgG1, IgG2a>IgG3>IgG2b). When mice were immunized with GST alone (100/50/50, 300 or 500 µg), antibodies against GST‐L1b or GST were not detected. However, GST doses of 10/10/10 or 200/100/100 µg induced an antibody response able to recognize both GST‐L1b and GST. We propose that an immunization protocol similar to the one used in this work may allow induction of high antibody titers specific against the parasite segment of a GST‐fusion protein.