Production, characterisation and use of monoclonal antibodies to human interleukin-5 in an enzyme-linked immunosorbent assay.
Two mouse monoclonal antibodies (Mabs) against recombinant human interleukin-5(rhIL-5) have been produced, characterised and purified. Both are IgG1 antibodies and neutralised the activity of rhIL-5 in the B13 assay. Neither Mab cross-reacted with mouse IL-5. A two-site sandwich enzyme-linked immunosorbent assay (ELISA) was developed with different combinations of the mouse Mabs and also a rat anti-mouse IL-5 Mab, TRFK5, which also has activity against rhIL-5. The most sensitive assay, with a lower detection limit of 0.5 ng/ml IL-5, used TRFK5 as the capture antibody and the mouse anti-human IL-5 Mab as second antibody. The sensitivity of this assay was increased by an enhanced chemiluminescent reagent and resulted in a lower limit of detection around 40 pg/ml IL-5.
McNamee
L A
LA
Division of Cellular and Molecular Sciences, Glaxo Group Research Ltd., Greenfold, Middx., U.K.
Fattah
D I
DI
Baker
T J
TJ
Bains
S K
SK
Hissey
P H
PH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D001710:Biotin; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D015848:Interleukin-5 / Q000032:analysis* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008808:Mice, Inbred CBA; D051381:Rats; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D015848:Interleukin-5; D001710:Biotin