Development and validation of an enzyme-linked immunosorbent assay (ELISA) for recombinant human gamma interferon.
A one-site ELISA for the quantification of recombinant human gamma interferon (rh-IFN-gamma) was developed and validated. A single monoclonal antibody (Mab) was used as a "catching" antibody and as a horseradish peroxidase (HRP)-labeled conjugate. Detection limit and quantification limit of this assay were estimated to be 1.26 and 15 ng/mL, respectively, and the coefficient of variation was below 15%. The ELISA was specific for rh-IFN-gamma, showing no cross reactivity to other related molecules in the range of the concentrations studied. The results correlated well with those obtained by a bioassay method. By using this assay, it was demonstrated that 0.01-1% (v/v) Tween 80 protected rh-IFN-gamma during freezing and thawing.
Bouyón
R
R
Center for Genetic Engineering and Biotechnology, Havana, Cuba. rebecca.bouyon@cigb.edu.cu
Santana
H
H
Pérez
E M
EM
Hernández
N
N
Furrazola
G
G
Abrahantes
M C
MC
D000911:Antibodies, Monoclonal / Q000276:immunology; D001666:Binding Sites, Antibody; D003429:Cross Reactions; D010450:Endopeptidases; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006358:Hot Temperature; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis* / Q000276:immunology; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D013501:Surface-Active Agents
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D013501:Surface-Active Agents; D007371:Interferon-gamma; D010450:Endopeptidases