Improvement in sensitivity of enzyme-linked immunosorbent assay for tumour necrosis factor.
Various protocols were used in the development of enzyme-linked immunosorbent assays (ELISA) to improve the sensitivity and range of detection of human tumour necrosis factor-alpha (TNF-alpha). ELISA can provide a specific, sensitive and rapid method for detection of TNF-alpha in patient's sera, and it is important that the assay used should be sufficiently sensitive to detect low levels of TNF-alpha. The double sandwich ELISA proved to be the most sensitive, detecting less than 0.080 ng/mL TNF. Of eight different protocols, one assay using a purified monoclonal antibody to human TNF-alpha and rabbit polyclonal anti-TNF-alpha antibody had the greatest sensitivity and range of detection. The study illustrates methods for the development of sensitive immunoassays which may have applications in many assay systems.
McLaughlin
P J
PJ
Department of Pathology, University of Melbourne, Parkville, Vic., Australia.
Elwood
N J
NJ
Ramadi
L T
LT
Pica
M R
MR
McKenzie
I F
IF
D000818:Animals; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007106:Immune Sera; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000032:analysis; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis*
D000911:Antibodies, Monoclonal; D007106:Immune Sera; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha