Increased frequency of both total and specific monoclonal antibody producing hybridomas using a fusion partner that constitutively expresses recombinant IL-6
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2022, Cell ReportsCitation Excerpt :Mice were boosted with 1 × 108 EID50 HKx31 i.p. on day 26. Two days later, spleens were harvested and 6 × 106 cells from each spleen were fused to an equal number of Sp2/mIL-6 hybridoma cells (ATCC® CRL-2016™) (Harris et al., 1992). The fusion was transferred to 96 or 384 well plates and incubated in HAT-(Sigma, St. Louis, MO) supplemented media (ClonaCell™-HY Medium E, Stemcell Technologies, Vancouver, BC) for ten days.
Generation of a panel of high affinity antibodies and development of a biosensor-based immunoassay for the detection of okadaic acid in shellfish
2015, ToxiconCitation Excerpt :The mouse with the highest affinity of the antiserum for its antigen, as determined by effective dose 50 (ED-50; the concentration of free OA required to inhibit the binding of the antiserum or antibody to the immobilised antigen by 50%), was selected for the generation of monoclonal antibodies by cellular fusion. Myeloma SP2/mIL-6 cells (Harris et al., 1992) were fused with spleen cells of the selected animal in the presence of polyethylene glycol (Köhler and Milstein, 1975). Individual clones were isolated by limiting dilution and antibody-producing clones were selected by their ability to bind and displace free OA on a competitive indirect immunoassay.
Predominant role for activation-induced cytidine deaminase in generating IgG anti-nucleosomal antibodies of murine SLE
2015, Journal of AutoimmunityCitation Excerpt :Monoclonal antibodies specific to the hapten p-azophenylarsonate (Ars) were generated by immunization of a B6 Aicda−/− mouse with 50 μg of Ars-kehyole limpet hemocyanin (KLH) i.p in alum at day 0, followed by 3 i.p. injections at days 7, 9 and 11 in PBS. At day 14, splenocytes were harvested and fused with SP2/0/mIL-6 cells [49]. Hybridoma culture supernatants were tested for the production of Igκ antibodies against chromatin or Ars-ovalbumin (OVA) as described above.
Enzymatic recycling-based amperometric immunosensor for the ultrasensitive detection of okadaic acid in shellfish
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