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dc.contributor.authorWaldron, E.E.
dc.contributor.authorMurray, Paul G.
dc.contributor.authorKolar, Zdenek
dc.contributor.authorYoung, Lawrence S.
dc.contributor.authorBrown, C.
dc.contributor.authorReynolds, Gary
dc.contributor.authorBaumforth, Karl R. N.
dc.contributor.authorToomey, S.
dc.contributor.authorAstley, S.J.
dc.contributor.authorPerera, Shantha
dc.contributor.authorNelson, Paul N.
dc.date.accessioned2008-06-19T16:14:34Z
dc.date.available2008-06-19T16:14:34Z
dc.date.issued2002
dc.identifier.citationHybridoma and Hybridomics, 21(5): 393-398
dc.identifier.issn1536-8599
dc.identifier.pmid12470483
dc.identifier.doi10.1089/153685902761022751
dc.identifier.urihttp://hdl.handle.net/2436/30219
dc.description.abstractThe 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.
dc.language.isoen
dc.publisherNew Rochelle (NY): Mary Ann Liebert, Inc.
dc.relation.urlhttp://www.liebertonline.com/doi/abs/10.1089%2F153685902761022751
dc.subjectMonoclonal antibodies
dc.subjectPeptides
dc.subjectMultiple Antigenic Peptides
dc.subject.meshAnimals
dc.subject.meshAntibodies, Monoclonal
dc.subject.meshBlotting, Western
dc.subject.meshEnzyme-Linked Immunosorbent Assay
dc.subject.meshHybridomas
dc.subject.meshImmunoglobulin Isotypes
dc.subject.meshMice
dc.subject.meshMice, Inbred BALB C
dc.subject.meshPeptides
dc.subject.meshSpleen
dc.subject.meshViral Matrix Proteins
dc.titleReactivity and isotype profiling of monoclonal antibodies using multiple antigenic peptides.
dc.typeJournal article
dc.identifier.journalHybridoma and Hybridomics
html.description.abstractThe 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.


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