A sandwich immunoassay to quantify low levels of turbot (Scophthalmus maximus) immunoglobulins.
We have developed an enzyme-linked immunosorbent assay for the quantification of turbot (Scophthalmus maximus) immunoglobulin (Ig). The capture antibody is a rabbit polyclonal antiserum to turbot Ig, and the detector antibody a monoclonal antibody (UR3) to the turbot Ig heavy chain. Both antibodies bind nearly 100% of turbot Ig. The assay allows detection of turbot Ig in serum at concentrations as low as 0.16 micrograms ml-1 and takes less than 4 h. Precision is satisfactory, with intra-assay coefficients of variation (CVs) ranging from 2.1 to 16.6%, and inter-assay CVs ranging from 5.8 to 24.6%. We used the assay to determine Ig concentrations in the sera of healthy turbot of different weights. Mean serum Ig concentration was 3.35 +/- 0.74 mg ml-1 for fish weighing 15-25 g and 11.14 +/- 1.87 mg ml-1 for fish weighing 1000-2000 g.
Estévez
J
J
Laboratorio de Parasitología, Facultad de Farmacia, Universidad de Santiago de Compostela, Santiago de Compostela, Spain.
Leiro
J
J
Santamarina
M T
MT
Ubeira
F M
FM
D000818:Animals; D000911:Antibodies, Monoclonal; D001835:Body Weight; D002850:Chromatography, Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005412:Flatfishes / Q000276:immunology*; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D007136:Immunoglobulins / Q000032:analysis* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests / Q000662:veterinary; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D007136:Immunoglobulins