Standardizing In Vitro ß-Lactam Antibiotic Allergy Testing with Synthetic IgE

Autores de IIS La Fe

• Ángel Maquieira Catalá

  Autor

• Sergi Morais Ezquerro

  Autor

Participantes ajenos a IIS La Fe

• Quintero-Campos, P
• Gozalbo-Rovira, R
• Rodríguez-Díaz, J

Grupos

• Unidad Mixta de Investigación en Nanomedicina y Sensores

Abstract

The global prevalence of & beta;-lactam allergy posesa major challengein administering first-line antibiotics, such as penicillins, to asignificant portion of the population. The lack of & beta;-lactamIgE antibody pools with defined selectivity hampers the standardizationand validation of in vitro assays for & beta;-lactam allergy testing.To address this limitation, this study introduces a synthetic IgEspecific to & beta;-lactam antibiotics as a valuable tool for drugallergy research and diagnostic tests. Using phage display technology,we constructed a library of human single-chain antibody fragments(scFv) to target the primary determinant of amoxicillin, a widelyused & beta;-lactam antibiotic. Subsequently, we produced a completehuman synthetic IgE molecule using the highly efficient baculovirusexpression vector system. This synthetic IgE molecule served as astandard in an in vitro chemiluminescence immunoassay for & beta;-lactamantibiotic allergy testing. Our results demonstrated a detection limitof 0.05 IU/mL (0.63 pM), excellent specificity (100%), and a four-foldhigher clinical sensitivity (73%) compared to the in vitro referenceassay when testing a cohort of 150 serum samples. These findings havesignificant implications for reliable interlaboratory comparison studies,accurate labeling of allergic patients, and combating the global publichealth threat of antimicrobial resistance. Furthermore, by servingas a valuable trueness control material, the synthetic IgE facilitatesthe standardization of diagnostic tests for & beta;-lactam allergyand demonstrates the potential of utilizing this synthetic strategyas a promising approach for generating reference materials in drugallergy research and diagnostics.