Development of monoclonal antibodies to gizzerosine, a toxic component present in fish meal.
This study is the first report of the development of monoclonal antibodies (MAbs) against gizzerosine (GZ), one of the causative agents of black vomit, a serious poultry disease. Balb/c mice were immunized with different GZ conjugates; the most immunogenic conjugate in experimental animals was determined by enzyme-linked immunoadsorbent assays (ELISA). Somatic fusions were carried out using splenic lymphocytes from GZ-immune mice and the NSO/2 myeloid cell line. Primary selection of hybridomas secreting antibodies to GZ was done using a direct ELISA, with GZ bound to bovine serum albumin (BSA), GZ directly bound to maleinimide preactivated plates and histamine bound to BSA, a GZ related biogenic amine present in fish meal. Four MAbs--3H4, 3H10, and 5B1 of the IgG1 isotype, and 8G7 of the Ig2a isotype-were specific to GZ and did not cross-react with histamine. Only monoclonals 3H4 and 8G7 bound GZ in solution by means of a competitive ELISA. Finally, to determine the performance of the competitive ELISA developed with the MAbs, experiments were conducted with GZ in solution (0 to 10 microg/ml) and with GZ labeled with horseradish peroxidase (HRP) as the tracer; the antibody complex was captured by using rabbit anti-mouse IgG preactivated ELISA plates. These experiments showed that monoclonal anti-GZ-3H4 generates a more sensitive assay close to linearity in the range about of 0.1 to 10 microg/ml of GZ. No cross-reaction was observed with histamine, histidine, or lysine at all concentrations tested.
Becker
M I
MI
Laboratorio de Inmunología, Departamento de Investigación y Desarrollo, BIOSONDA S.A., Santiago, Chile.
Carrasco
I
I
Beltran
C
C
Torres
M
M
Jaureguiberry
B
B
De Ioannes
A E
AE
D000821:Animal Feed / Q000032:analysis; D000911:Antibodies, Monoclonal; D001679:Biogenic Amines / Q000032:analysis* / Q000276:immunology / Q000633:toxicity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005394:Fish Flour / Q000032:analysis; D006825:Hybridomas; D007093:Imidazoles / Q000032:analysis* / Q000276:immunology / Q000633:toxicity; D014118:Toxins, Biological / Q000032:analysis
D000911:Antibodies, Monoclonal; D001679:Biogenic Amines; D007093:Imidazoles; D014118:Toxins, Biological; C045114:gizzerosine