Article Text
Abstract
Background: Two mouse monoclonal antibodies have been described, namely: mAb 2C7 (IgG2bκ), which is directed against the major house dust mite allergen Der p 1, and mAb 2G10 (IgG1κ), which is an anti-idiotypic antibody raised against mAb 2C7. Given its broad IgE specificity, anti-idiotype mAb 2G10 could potentially have immunomodulatory applications. For example, a chimaeric human IgG version of mAb 2G10 could prove to be a useful molecule for binding to mast cell and basophil FcεRI bound IgE, and in doing so co-ligating FcεRI with FcγRIIB, which has been reported to have downregulatory effects.
Aims: To produce a chimaeric human IgE version of mAb 2C7 (mAb 2C7huE) and a chimaeric human IgG1 version of its anti-idiotype mAb 2G10 (mAb 2G10huG1).
Methods: The Vκ and VH regions of mAb 2C7 and its anti-idiotype mAb 2G10 were engineered into human constant regions of the IgE and IgG1 isotypes, respectively.
Results: The production of chimaeric mAb 2C7huE and its anti-idiotype mAb 2G10huG1 confirmed that the respective mouse antibody V regions were successfully engineered into human constant regions and still retained the specificity of the original murine V regions.
Conclusion: The newly constructed chimaeric antibodies will be useful to investigate the downregulation of IgE mediated hypersensitivity by the crosslinking of FcεRI with FcγRIIB.
- antibody
- anti-idiotype
- chimaerisation
- Der 1
- IgE
- CDR, complementarity determining region
- ELISA, enzyme linked immunosorbent assay
- FcεRI, high affinity IgE receptor
- FcγRIIB, low affinity IgE receptor
- FCS, fetal calf serum
- FRW, framework region
- Ig, immunoglobulin
- IMDM, Iscove’s modified Dulbecco’s medium
- ITAM, immunoreceptor tyrosine based activation motif
- mAb, monoclonal antibody
- PBS, phosphate buffered saline
- PCR, polymerase chain reaction
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- CDR, complementarity determining region
- ELISA, enzyme linked immunosorbent assay
- FcεRI, high affinity IgE receptor
- FcγRIIB, low affinity IgE receptor
- FCS, fetal calf serum
- FRW, framework region
- Ig, immunoglobulin
- IMDM, Iscove’s modified Dulbecco’s medium
- ITAM, immunoreceptor tyrosine based activation motif
- mAb, monoclonal antibody
- PBS, phosphate buffered saline
- PCR, polymerase chain reaction