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Primary sequence and molecular model of the variable region of a mouse monoclonal anti‐Der p 1 antibody showing a similar epitope specificity as human IgE
Author(s) -
McElveen Je,
Mike Clark,
S. J. Smith,
Sewell Hf,
F Shakib
Publication year - 1998
Publication title -
clinical and experimental allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 154
eISSN - 1365-2222
pISSN - 0954-7894
DOI - 10.1046/j.1365-2222.1998.00378.x
Subject(s) - immunoglobulin e , monoclonal antibody , epitope , microbiology and biotechnology , immunoglobulin light chain , biology , antibody , isotype , complementary dna , house dust mite , allergen , homology (biology) , peptide sequence , immunology , allergy , gene , genetics
Background Der p 1, a major mite allergen, elicits IgE antibody responses in 80% of patients suffering from dust mite allergy. Given the potent IgE eliciting properties of Der p 1, there is considerable interest in studying the molecular architecture of the variable (Fv) region of IgE antibodies specific for this allergen. Objectives IgE is present in human serum at extremely low concentrations, and as such it is practically impossible to purify sufficient quantities for structural studies. We have therefore sought to sequence and model a representative murine monoclonal (MoAb) anti‐Der p 1 antibody, as a surrogate human IgE. Methods The cDNA coding for the Fv region of an anti‐Der p 1 MoAb (2C7), that mimics the binding of human IgE to Der p 1, was amplified by PCR, cloned and sequenced. The predicted amino acid sequences were then compared with a directory of human germline V‐gene segments. Modelling of the Fv region of MoAb 2C7 was carried out using the extensive database of existing immunoglobulin structures in the Brookhaven PDB. Results The MoAb 2C7 heavy chain showed greater than 70% homology with three members of the V H 3 family, DP‐35, DP‐53 and DP‐54. Similarly, the light chain showed greater than 70% homology with 11 V K sequences, including the V K II sequences DPK18, DPK19 and DPK28. A molecular model of the Fv region of MoAb 2C7 was generated and can be accessed from the EMBL databank. Conclusions Antibodies similar to MoAb 2C7 could be generated as part of the human repertoire. The availability of 3‐dimensional model of MoAb 2C7, as a surrogate human IgE antibody, combined with further data on its epitope specificity, will facilitate studies into IgE antibody responses to Der p 1.