Cytokine-deficient mice as a model for generation of autologous anti-cytokine monoclonal antibodies

Shichkin, Valentin P.; Spivak, N. Ya.

doi:10.1016/j.imlet.2005.08.005

Cited by 2 publications

(1 citation statement)

References 36 publications

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“…With this strategy monoclonal antibodies against the prion protein (the infective agent of scrapie) was possible [12]. This method was also successfully used to make a monoclonal antibody against apolipoproteins [13], while another group claimed successful generation of polyclonal but failed in generation of monoclonal antibody against cytokines [14]. We have also successfully used the described strategy to obtain an antibody against another related protein – mouse acetylcholinesterase (data not shown).…”

Section: Discussionmentioning

confidence: 96%

A Novel System for the Efficient Generation of Antibodies Following Immunization of Unique Knockout Mouse Strains

2010

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BackgroundWe wished to develop alternate production strategies to generate antibodies against traditionally problematic antigens. As a model we chose butyrylcholinesterase (BChE), involved in termination of cholinergic signaling, and widely considered as a poor immunogen.Methodology/Principal FindingsJettisoning traditional laborious in silico searching methods to define putative epitopes, we simply immunized available BChE knock-out mice with full-length recombinant BChE protein (having been produced for crystallographic analysis). Immunization with BChE, in practically any form (recombinant human or mouse BChE, BChE purified from human serum, native or denatured), resulted in strong immune responses. Native BChE produced antibodies that favored ELISA and immunostaining detection. Denatured and reduced BChE were more selective for antibodies specific in Western blots. Two especially sensitive monoclonal antibodies were found capable of detecting 0.25 ng of BChE within one min by ELISA. One is specific for human BChE; the other cross-reacts with mouse and rat BChE. Immunization of wild-type mice served as negative controls.Conclusions/SignificanceWe examined a simple, fast, and highly efficient strategy to produce antibodies by mining two expanding databases: namely those of knock-out mice and 3D crystallographic protein-structure analysis. We conclude that the immunization of knock-out mice should be a strategy of choice for antibody production.

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Section: Discussionmentioning

confidence: 96%

A Novel System for the Efficient Generation of Antibodies Following Immunization of Unique Knockout Mouse Strains

2010

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Properties of Hybridomas Generated by Spleen Cells of IL-4– and IL-13–Knockout Mice

Shichkin

Spivak

2005

Hybridoma

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Interleukin-4 (IL-4)- and IL-13-knockout mice were immunized with murine recombinant IL-4 or IL-13, and spleen cells were fused with P3X63-Ag8.653 myeloma cells. Selection of the antigen-positive hybridomas was fulfilled in the presence of IL-6 containing thymic stroma cell supernatant (TSS). All of the selected anti- IL-4- and anti-IL-13-specific hybridoma clones (eight and 10, respectively) required the presence of TSS (0.5-2.5%) for their cloning, stable growth in large-scale cultures, and production of monoclonal antibodies (MAbs). Several of the anti-IL-4-specific clones were adapted to growth without TSS. However, the loss of antibody-secreting capacity in the process of adaptation to TSS-free growth was detected. The data demonstrate that cytokine-deficient mice technology can be used for generation of MAbs to autologous cytokines.

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Cytokine-deficient mice as a model for generation of autologous anti-cytokine monoclonal antibodies

Cited by 2 publications

References 36 publications

A Novel System for the Efficient Generation of Antibodies Following Immunization of Unique Knockout Mouse Strains

A Novel System for the Efficient Generation of Antibodies Following Immunization of Unique Knockout Mouse Strains

Properties of Hybridomas Generated by Spleen Cells of IL-4– and IL-13–Knockout Mice

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