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dc.contributor.authorMeager, A
dc.contributor.authorParti, S
dc.contributor.authorLeung, H
dc.contributor.authorWoolley, J
dc.contributor.authorPeil, E
dc.contributor.authorSidhu, S
dc.contributor.authorRoberts, Trudy
dc.date.accessioned2010-11-08T10:36:06Z
dc.date.available2010-11-08T10:36:06Z
dc.date.issued1987-11-23
dc.identifier.citationA two-site sandwich immunoradiometric assay of human lymphotoxin with monoclonal antibodies and its applications. 1987, 104 (1-2):31-42 J. Immunol. Methodsen
dc.identifier.issn0022-1759
dc.identifier.pmid3119725
dc.identifier.doi10.1016/0022-1759(87)90484-4
dc.identifier.urihttp://hdl.handle.net/10541/114926
dc.description.abstractThree monoclonal antibodies (MoAbs L49-15, L81-11 and L238-14) were raised against recombinant human lymphotoxin (rLT) derived from E. coli containing the cDNA sequence specifying LT. MoAb L81-11 strongly neutralised the cytotoxicity of LT derived either from E. coli or the RPMI 1788 lymphoblastoid cell line, whilst the other two MoAbs were only weakly neutralising in this respect. L81-11 and L238-14 MoAbs bound to different antigenic determinants on the rLT molecule, but neither bound to other lymphokines such as the structurally related tumour necrosis factor (TNF). As such, these MoAbs were ideal reagents for immunoassay of LT and a very sensitive, highly specific immunoradiometric assay (IRMA) was developed. This assay was rapid to perform and was capable of detecting as little as 10 pg/ml of LT. Application of the LT IRMA in combination with previously developed human gamma-interferon (IFN-gamma) and human TNF-specific IRMA (Crane et al., 1985; Meager et al., 1987) permitted independent estimations of these three substances to be carried out in parallel. By these means, it was found that RPMI 1788 produced both LT and TNF, but not IFN-gamma. Extensive analyses on cytokine (monokine and lymphokine) preparations derived from a variety of activated lymphocytes are also reported. Co-production of LT, TNF and IFN-gamma was a common finding, even occurring in alloantigen-specific T helper cell clones.
dc.language.isoenen
dc.subjectTumour Necrosis Factor-alphaen
dc.subject.meshAntibodies, Monoclonal
dc.subject.meshBiological Products
dc.subject.meshChromatography
dc.subject.meshCytokines
dc.subject.meshCytotoxins
dc.subject.meshHumans
dc.subject.meshInterferon-gamma
dc.subject.meshLymphocyte Activation
dc.subject.meshLymphocytes
dc.subject.meshLymphotoxin-alpha
dc.subject.meshRadioimmunoassay
dc.subject.meshTumor Necrosis Factor-alpha
dc.titleA two-site sandwich immunoradiometric assay of human lymphotoxin with monoclonal antibodies and its applications.en
dc.typeArticleen
dc.contributor.departmentNational Institute for Biological Standards and Control, Potters Bar, Herts, U.K.en
dc.identifier.journalJournal of Immunological Methodsen
html.description.abstractThree monoclonal antibodies (MoAbs L49-15, L81-11 and L238-14) were raised against recombinant human lymphotoxin (rLT) derived from E. coli containing the cDNA sequence specifying LT. MoAb L81-11 strongly neutralised the cytotoxicity of LT derived either from E. coli or the RPMI 1788 lymphoblastoid cell line, whilst the other two MoAbs were only weakly neutralising in this respect. L81-11 and L238-14 MoAbs bound to different antigenic determinants on the rLT molecule, but neither bound to other lymphokines such as the structurally related tumour necrosis factor (TNF). As such, these MoAbs were ideal reagents for immunoassay of LT and a very sensitive, highly specific immunoradiometric assay (IRMA) was developed. This assay was rapid to perform and was capable of detecting as little as 10 pg/ml of LT. Application of the LT IRMA in combination with previously developed human gamma-interferon (IFN-gamma) and human TNF-specific IRMA (Crane et al., 1985; Meager et al., 1987) permitted independent estimations of these three substances to be carried out in parallel. By these means, it was found that RPMI 1788 produced both LT and TNF, but not IFN-gamma. Extensive analyses on cytokine (monokine and lymphokine) preparations derived from a variety of activated lymphocytes are also reported. Co-production of LT, TNF and IFN-gamma was a common finding, even occurring in alloantigen-specific T helper cell clones.


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