Recombinant Phleum pratense Pollen allergen Phl p 5a is produced in E. coli with a partial protein length of 1-286 amino acids. The protein carries an N-terminal 10xHis-SUMO tag along with a C-terminal Myc tag, which helps with purification and detection processes. SDS-PAGE analysis shows the product achieves greater than 85% purity, suggesting it meets quality standards for research applications. This product is intended for research use only.
Phleum pratense Pollen allergen Phl p 5a appears to be a significant component when studying grass pollen allergies. As an allergenic protein, it likely plays a crucial role in immune responses that pollen exposure triggers. Research into Phl p 5a tends to focus on understanding how it interacts with the immune system. This work may contribute to developing allergy diagnostics and potential therapeutic interventions.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
As a pollen allergen, Phl p 5a's bioactivity depends on correct conformational epitopes for IgE antibody binding. E. coli expression typically cannot replicate eukaryotic post-translational modifications (e.g., glycosylation) that may affect allergenicity. The large dual-tag system (especially the N-terminal SUMO tag) may sterically hinder proper folding of conformational epitopes. While the protein may contain linear epitopes, its ability to mimic native allergen structure is uncertain without validation. High purity reduces non-specific effects but doesn't guarantee native folding. Therefore, this protein is unlikely to possess complete bioactivity (IgE binding capacity) without experimental confirmation.
1. Allergen-Specific Antibody Development and Characterization
This recombinant Phl p 5a can serve as an immunogen for antibody development targeting linear epitopes. The dual tags facilitate purification and detection during screening. However, antibodies generated may not recognize conformational epitopes of natural Phl p 5a due to potential misfolding. Validate antibody binding against native allergen extracts from timothy grass pollen for immunological applications.
2. Protein-Protein Interaction Studies Using Pull-Down Assays
The His tag enables immobilization for pull-down assays, but results may not reflect physiological interactions if the protein is misfolded. The tags themselves may cause non-specific binding. Any identified interactions must be confirmed using natural Phl p 5a to ensure biological relevance, especially for studies involving immune receptors like IgE.
3. Structural and Biochemical Characterization Studies
This protein is suitable for basic biophysical analysis (e.g., circular dichroism for secondary structure, dynamic light scattering for aggregation state). However, structural conclusions may not reflect the native allergen due to potential tag-induced conformational changes. Remove tags for meaningful structural insights into natural Phl p 5a.
4. Cross-Reactivity Analysis in Allergen Research
This protein can be used for cross-reactivity studies only if validated to retain conformational epitopes. Otherwise, linear epitope cross-reactivity may be detected, but this doesn't fully represent clinical allergen cross-reactivity. Always compare results with natural allergen extracts and include IgE binding assays from allergic patient sera for clinical relevance.
Final Recommendation & Action Plan
Before using this recombinant Phl p 5a for functional studies, validate its structural integrity and allergenicity. Perform circular dichroism to compare secondary structure with natural Phl p 5a, and conduct IgE binding ELISAs using sera from timothy grass-allergic patients to confirm antigenicity. If validation fails, limit use to linear epitope mapping or antibody production against linear sequences. For reliable allergen research, consider eukaryotic expression systems (e.g., insect cells) that better replicate natural protein folding and modifications. Always include natural Phl p 5a as a positive control in experiments.
