The use of a double antibody sandwich ELISA and monoclonal antibodies for the assessment of porcine IgM, IgG and IgA concentrations.
Double antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) have been developed for the assessment of IgM, IgG and IgA concentrations in porcine serum. Isotype-specific monoclonal antibodies (mAb) were used for coating and detection. The DAS-ELISAs were examined for their ability to detect each isotype and for the assay variations. A computer programme was used to verify the parallelism of the slope of each serum sample with the slope of standard/reference serum, a prerequisite for reliable estimation of Ig concentrations. The DAS-ELISAs are easy to perform and highly specific, have adequate detection levels (ranging from 7 to 50 ng ml-1) and are very reproducible, as illustrated by the inter- and intra-assay variation coefficients (ranging between 4.9 and 7%). To illustrate the applicability of the ELISAs we assessed Ig concentrations in pig sera sampled from birth to young adulthood.
Bianchi
A T
AT
Department of Immunology, Central Veterinary Institute, Lelystad, Netherlands.
Moonen-Leusen
H W
HW
van der Heijden
P J
PJ
Bokhout
B A
BA
D000375:Aging / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D007070:Immunoglobulin A / Q000032:analysis*; D007074:Immunoglobulin G / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M