ORIGINAL ARTICLE

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

P B Furtado¹, J E McElveen¹, L Gough¹, K L Armour², M R Clark², H F Sewell¹ and F Shakib¹

¹ Division of Molecular and Clinical Immunology, University of Nottingham, Faculty of Medicine and Health Sciences, Queen’s Medical Centre, Nottingham NG7 2UH, UK
² Immunology Division, Department of Pathology, Cambridge University, Tennis Court Road, Cambridge CB2 1QP, UK

Correspondence to:
Correspondence to:
Dr F Shakib, Division of Molecular and Clinical Immunology, Faculty of Medicine and Health Sciences, Queen’s Medical Centre, Nottingham NG7 2UH, UK;
farouk.shakib{at}nottingham.ac.uk

Background: Two mouse monoclonal antibodies have been described, namely: mAb 2C7 (IgG2b), which is directed against the major house dust mite allergen Der p 1, and mAb 2G10 (IgG1), 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 FcRI bound IgE, and in doing so co-ligating FcRI with FcRIIB, 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 V 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 FcRI with FcRIIB.

Keywords: antibody; anti-idiotype; chimaerisation; Der 1; IgE

Abbreviations: CDR, complementarity determining region; ELISA, enzyme linked immunosorbent assay; FcRI, high affinity IgE receptor; FcRIIB, 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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