The production and characterisation of a chimaeric human IgE antibody, recognising the major mite allergen Der p 1, and its chimaeric human IgG1 anti-idiotype. Issue 5 (1st October 2002)

• Record Type: Journal Article
• Authors: Furtado, P B; McElveen, J E; Gough, L; Armour, K L; Clark, M R; Sewell, H F; Shakib, F
• Is Part Of: Journal of clinical pathology. Volume 55:Issue 5(2002)
• Page Start: 315
• Page End: 324
• Publication Date: 2002-10-01
• Subjects: 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; Pathology -- Periodicals; Pathology, Molecular -- Periodicals; 616.0705
• DOI: 10.1136/mp.55.5.315
• Languages: English
• ISSNs: 0021-9746

Abstract

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.