Article Text

Download PDFPDF
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
  1. P B Furtado1,
  2. J E McElveen1,
  3. L Gough1,
  4. K L Armour2,
  5. M R Clark2,
  6. H F Sewell1,
  7. F Shakib1
  1. 1Division of Molecular and Clinical Immunology, University of Nottingham, Faculty of Medicine and Health Sciences, Queen’s Medical Centre, Nottingham NG7 2UH, UK
  2. 2Immunology Division, Department of Pathology, Cambridge University, Tennis Court Road, Cambridge CB2 1QP, UK
  1. 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

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

Statistics from Altmetric.com

Request Permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.

Footnotes