| Title |
Evaluation of inhibitory activity of novel monoclonal antibodies against cat allergen Fel d 7 and their application to analyse allergen extracts |
| Authors |
Rudokas, Vytautas ; Pena-Amelunxen, Glorismer ; Briza, Peter ; Aglas, Lorenz ; Žvirblienė, Aurelija |
| DOI |
10.1111/sji.70056 |
| Full Text |
|
| Is Part of |
Scandinavian journal of immunology.. John Wiley & Sons Ltd.. 2025, vol. 102, iss. 3, art. no. e70056, p. [1-12].. ISSN 0300-9475. eISSN 1365-3083 |
| Keywords [eng] |
allergen extract ; allergy ; cat ; Fel d 7 ; Felis domesticus ; monoclonal antibody |
| Abstract [eng] |
Current characterisation of cat dander/hair extracts used for allergy diagnosis or allergen-specific immunotherapy is mainly standardised towards the major allergen Fel d 1, while other allergens such as the lipocalin Fel d 7 are insufficiently characterised in such allergen products. This study aimed to produce recombinant Fel d 7 (rFel d 7) and murine IgG monoclonal antibodies (mAbs) specific to it for quantification in allergen extracts and assess the potential of the mAbs in inhibiting patients' IgE in functional assays. rFel d 7 was expressed in E. coli, purified by Ni-affinity and ion exchange chromatography, and physicochemically characterised by circular dichroism, Fourier-transform infrared spectroscopy, dynamic light scattering and mass spectrometry. Twenty hybridoma cell lines producing Fel d 7-specific mAbs were generated and sandwich ELISA was established for the quantitation of Fel d 7. Six different cat allergen extracts from different manufacturers and prepared from different sources were analysed and the concentration ranged from 0.02 μg/mg to 22.59 μg/mg. A mAb pool recognising non-overlapping epitopes inhibited the binding of human IgE-antigen complex formation (63.7% highest inhibition) and IgE-Fel d 7 cross-linking and consequent degranulation of basophilic cells (57.2% highest inhibition). We demonstrate the vast difference of Fel d 7 content in different cat allergen extracts, highlighting the necessity of improved standardisation of cat allergen extracts. The inhibition results showed that the analysed mAbs effectively inhibit rFel d 7 binding to human IgE, an assay we recommend for assessing the potency of allergen extracts as part of extract standardisation. |
| Published |
John Wiley & Sons Ltd |
| Type |
Journal article |
| Language |
English |
| Publication date |
2025 |
| CC license |
|
