Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico, Universitat Politècnica de València-Universitat de València, Camino de vera s/n, 46022, Valencia, Spain.
Departamento de Química, Universitat Politècnica de València, Camino de Vera s/n, 46022, Valencia, Spain.
Sci Rep. 2020 Sep 29;10(1):16037. doi: 10.1038/s41598-020-73109-w.
New antigens deriving from -lloyl and -llanyl, major and minor determinants, respectively, were produced for β-lactam antibiotics cefuroxime, cefotaxime, ceftriaxone, meropenem and aztreonam. Twenty β-lactam antigens were produced using human serum albumin and histone H1 as carrier proteins. Antigens were tested by multiplex in vitro immunoassays and evaluated based on the detection of specific IgG and IgE in the serum samples. Both major and minor determinants were appropriate antigens for detecting specific anti-β-lactam IgG in immunised rabbit sera. In a cohort of 37 allergic patients, we observed that only the minor determinants (-llanyl antigens) were suitable for determining specific anti-β-lactam IgE antibodies with high sensitivity (< 0.01 IU/mL; 24 ng/L) and specificity (100%). These findings reveal that not only the haptenisation of β-lactam antibiotics renders improved molecular recognition events when the 4-member β-lactam ring remains unmodified, but also may contribute to develop promising minor antigens suitable for detecting specific IgE-mediated allergic reactions. This will facilitate the development of sensitive and selective multiplexed in vitro tests for drug-allergy diagnoses to antibiotics cephalosporin, carbapenem and monobactam.
针对β-内酰胺抗生素头孢呋辛、头孢噻肟、头孢曲松、美罗培南和氨曲南,分别产生了源自-lloyl 和 -llanyl 的新抗原,分别为主要和次要决定簇。使用人血清白蛋白和组蛋白 H1 作为载体蛋白,共制备了 20 种β-内酰胺抗原。通过多重体外免疫测定法对这些抗原进行了测试,并根据血清样本中特异性 IgG 和 IgE 的检测结果进行了评估。主要和次要决定簇均适合检测免疫兔血清中的特异性抗β-内酰胺 IgG。在 37 名过敏患者的队列中,我们观察到只有次要决定簇(-llanyl 抗原)适合检测特异性抗β-内酰胺 IgE 抗体,具有高灵敏度(<0.01 IU/mL;24 ng/L)和特异性(100%)。这些发现表明,不仅β-内酰胺抗生素的半抗原化在 4 元β-内酰胺环保持不变的情况下会导致改善的分子识别事件,而且还可能有助于开发合适的次要抗原,用于检测特异性 IgE 介导的过敏反应。这将有助于开发针对头孢菌素、碳青霉烯和单酰胺类抗生素的药物过敏诊断的敏感和选择性多重体外检测。