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    A two-site sandwich immunoradiometric assay of human lymphotoxin with monoclonal antibodies and its applications.

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    Authors
    Meager, A
    Parti, S
    Leung, H
    Woolley, J
    Peil, E
    Sidhu, S
    Roberts, Trudy
    Affiliation
    National Institute for Biological Standards and Control, Potters Bar, Herts, U.K.
    Issue Date
    1987-11-23
    
    Metadata
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    Abstract
    Three 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.
    Citation
    A two-site sandwich immunoradiometric assay of human lymphotoxin with monoclonal antibodies and its applications. 1987, 104 (1-2):31-42 J. Immunol. Methods
    Journal
    Journal of Immunological Methods
    URI
    http://hdl.handle.net/10541/114926
    DOI
    10.1016/0022-1759(87)90484-4
    PubMed ID
    3119725
    Type
    Article
    Language
    en
    ISSN
    0022-1759
    ae974a485f413a2113503eed53cd6c53
    10.1016/0022-1759(87)90484-4
    Scopus Count
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    All Paterson Institute for Cancer Research

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