LETI Pharma SLU

A functional allergen quantification kit requires two key components: a pair of monoclonal antibodies (mAbs) that do not interfere with each other at the protein binding site, and a reference standard with sufficient purity and stability. Despite the clinical relevance of Jun a 1 in patients with Cupressaceae pollinosis, no commercial kits are currently available for its quantification in allergenic extracts. The aim of this study was to develop and validate a sandwich ELISA kit for the quantification of Jun a 1 in Juniperus ashei extracts, using appropriately selected mAbs and a stable standard. To this end, Jun a 1 was purified to >95% purity, as confirmed by SDS-PAGE, Western blot, and mass spectrometry analysis. Its stability was demonstrated after 48 months of storage. The linear B-cell epitopes recognized by the mAbs were identified and mapped onto the Jun a 1 structure, confirming that they are non-overlapping. Following validation, the ELISA method proved to be specific, linear, accurate, and precise. In conclusion, we successfully developed and validated an ELISA assay for the quantification of Jun a 1 in J. ashei extracts, supported by the generation of suitable monoclonal antibodies and a stable reference standard.

Limited evidence indicates that baked milk (BM) intake accelerates cow's milk allergy (CMA) resolution or modulates immunity in BM tolerant (BMT) and BM-allergic (BMA) phenotypes.

This study evaluates the efficacy, safety, and immunological effects of BM introduction, focusing on regular intake in BMT and low-dose intake in BMA, and their impact on tolerance development.

A prospective study including children 12 to 72 months of age with an oral food challenge (OFC) confirmed CMA. Children with CMA were categorized into BMT or BMA phenotypes based on BM-OFC results. BMT children were randomized into 2 groups: B2 (daily intake of 0.55 g of BM protein [BMP]) and C2 (strict avoidance of BM). BMA children who tolerated the minimal dose were similarly randomized: B1 (daily intake of 0.0375 g of BMP) and C1 (strict avoidance of BM). Assessments at baseline (T0) and 12 months (T1) included skin prick tests (SPT), specific-IgE (sIgE), specific-IgG4 (sIgG4), basophil activation test (BAT), and T-cell cytokine profile. BM doses doubled at 6 months, and CM-OFC confirmed outcomes at T1.

A total of 50 participants with confirmed CMA were included, of whom 32 were BMT and 18 BMA. Daily BM intake in the B2 group significantly enhanced tolerance to CM (77.2% vs 40% in C2, P < .05), with improved safety and marked reductions in SPT and sIgE markers. Reactivity thresholds increased 9-fold (0.9 g vs 0.05 g). No significant increase in tolerance was seen in the B1 group. Mild home reactions occurred in 9.3%. Predictors of tolerance included early age (<24 months), BM consumption, absence of asthma, immunological markers (sIgE, SPT, and sIgG4/sIgE), and reduced BAT reactivity.

Early BM introduction appears to support CMA resolution in BMT and may enhance safety, particularly in young children. Regular BM consumption could contribute to better outcomes, while biomarkers offer valuable insights for personalized management.