The ZeBaP project, funded by the German Federal Ministry of Education and Research (grant 281A301D18) and carried out from 1 April 2020 to 31 August 2023, aimed to create a cell‑based assay that could quantify the allergenic potential of processed foods and to develop a new extraction buffer that would improve the recovery of allergenic proteins from such matrices. The core technical outcome was the formulation of a novel, environmentally friendly extraction medium that outperformed the previously used buffer in both SDS‑PAGE and ELISA analyses. When applied to baked cookies and chocolate spiked with peanut protein, the new buffer produced a markedly stronger protein band on SDS‑PAGE and a higher recovery rate in ELISA, indicating that the solubility of allergenic proteins was significantly enhanced. The buffer’s composition was optimized by testing a range of pH values and buffering agents; the final recipe maintained antibody binding at neutral to slightly alkaline pH while avoiding harsh chemicals that could damage cell cultures. In addition, the project investigated how roasting degree affected peanut and hazelnut allergen solubility. Chemical analyses of nuts roasted to different temperatures revealed that higher roasting levels reduced the extractability of allergenic proteins, a finding that underscores the need for robust extraction methods in processed food testing.

Although the cell‑based assay was designed to use a humanised basophilic cell line derived from the Paul‑Ehrlich‑Institut, the project could not complete this part because the cell line could not be modified to the required extent. Consequently, the assay did not provide a viable alternative to conventional ELISA or PCR methods, and the focus shifted entirely to the buffer development. The new extraction medium is now slated for commercial release in a test kit that will become available in May 2024. R‑Biopharm AG, the project lead, plans to integrate the buffer into an automated extraction platform, thereby offering a multi‑parameter solution for allergen detection that is both user‑friendly and compliant with environmental regulations.

Collaboration was central to the project’s progress. R‑Biopharm AG coordinated the overall effort, overseeing experimental design, data analysis, and project reporting. The Technical University of Dresden contributed expertise in the chemical characterization of roasted nuts and provided model food matrices for testing. The Paul‑Ehrlich‑Institut supplied the basophilic cell line and assisted in assay validation, while RefLab, a subsidiary of R‑Biopharm, is positioned to commercialise the cell‑based testing approach once the technical hurdles are resolved. The project’s timeline included a reporting period from 1 April 2022 to 31 August 2023, during which the new buffer was finalized and validated. No peer‑reviewed publications have yet been released, as the extraction buffer remains proprietary and is intended for exclusive use in the forthcoming commercial kit.