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Microarray-based evaluation of selected recombinant timothy grass allergens expressed in E. Coli and N. Benthamiana
Laimis Silimavicius1,2, Lieve Tchebotarev3, Mindaugas Zaveckas4
1Institute of Biotechnology, Life Sciences Center, Vilnius University, Sauletekio av. 7, Vilnius, 10257, Lithuania. laimis.silimavicius@gmc.vu.lt.
BMC Biotechnology
|October 4, 2024
Summary
Recombinant timothy grass allergens produced in E. coli and N. benthamiana show diagnostic potential on microarrays. Phl p 11 (MBP-fused) demonstrated high specificity and sensitivity, though intra-assay variation requires optimization.
Area of Science:
- Allergen immunotherapy
- Molecular biology
- Diagnostic assay development
Background:
- Timothy grass (Phleum pratense) is a major allergen source.
- Recombinant allergens are increasingly used for allergy diagnostics.
- Optimizing production systems for diagnostic accuracy is crucial.
Purpose of the Study:
- To compare diagnostic performance of recombinant timothy grass allergens.
- To evaluate allergens produced in E. coli and N. benthamiana.
- To utilize a custom-made microarray chip for analysis.
Main Methods:
- Produced recombinant timothy grass allergens (Phl p 1-6, 11, 12) in E. coli and N. benthamiana.
- Tested 113 patient serum samples on a custom microarray.
- Assessed diagnostic sensitivity, specificity, and IgE correlation with commercial tests (ALEX, ISAC).
Main Results:
- Phl p 1, 2, 5, 6, and 11 showed high positive correlation with commercial tests.
- MBP-fused Phl p 11 exhibited high diagnostic specificity and sensitivity (0.3 cut-off).
- Significant intra-assay variation was observed for some allergens.
Conclusions:
- Recombinant allergens from E. coli and N. benthamiana show diagnostic promise on microarrays.
- MBP-fused Phl p 11 is a high-performing diagnostic marker.
- Further optimization of allergen formulation and storage is needed to address intra-assay variation.
