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    Reactivity and isotype profiling of monoclonal antibodies using multiple antigenic peptides.

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    Authors
    Waldron, E.E.
    Murray, Paul G.
    Kolar, Zdenek
    Young, Lawrence S.
    Brown, C.
    Reynolds, Gary
    Baumforth, Karl R. N.
    Toomey, S.
    Astley, S.J.
    Perera, Shantha
    Nelson, Paul N.
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    Issue Date
    2002
    
    Metadata
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    Abstract
    The characterisation of monoclonal antibodies (MAbs) is essential for the development of assay systems particularly where antigens have been developed using synthetic peptides. Indeed some peptide-carrier conjugates fail to induce immune responses and may not generate antibodies that bind to native protein. As an alternative to peptide-carrier conjugates, multiple antigenic peptides (MAPs) have been used for immunization strategies, but with little regard to the characteristics of the MAbs produced. In this study, we used 3 MAPs of Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) to immunise BALB/c mice. Overall, the polyclonal antibody responses from tail bleeds showed that MAPs evoked B-cell responses. However, on screening 144 hybridomas, 24 MAb supernatants exhibited weak to moderate reactivity in enzyme-linked immunosorbant assay (ELISA) and against cell cytospin preparations (B95.8 and AG876 LCL), respectively. Isotype profiling of hybridoma supernatants also showed that 11 out of 24 were IgM. Further characterization of 6 MAbs in Western blotting showed reactivity to recombinant LMP1 and only one MAb (B28D) showed weak reactivity to the malignant cells (Hodgkin/Reed-Sternberg; HRS cells) of an EBV+ Hodgkin's lymphoma using paraffin-embedded tissue. It is probable that these MAPs failed to augment T-cell help and contributed to the production of low affinity (IgM) antibodies. These observations may be of importance to future immunization strategies, where MAPs are used in the production of monoclonal reagents.
    Citation
    Hybridoma and Hybridomics, 21(5): 393-398
    Publisher
    New Rochelle (NY): Mary Ann Liebert, Inc.
    Journal
    Hybridoma and Hybridomics
    URI
    http://hdl.handle.net/2436/30219
    DOI
    10.1089/153685902761022751
    PubMed ID
    12470483
    Additional Links
    http://www.liebertonline.com/doi/abs/10.1089%2F153685902761022751
    Type
    Journal article
    Language
    en
    ISSN
    1536-8599
    ae974a485f413a2113503eed53cd6c53
    10.1089/153685902761022751
    Scopus Count
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    Research Institute in Healthcare Science

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