Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic.

摘要: Allergic reactions towards β-lactam antibiotics pose an important clinical problem. The ability of small molecules, such as a β-lactams, to bind covalently proteins, in process known haptenation, is considered necessary for induction specific immunological response. Identification the proteins modified by β-lactams and elucidation relevance this allergic requires sensitive tools. Here we describe preparation characterization biotinylated amoxicillin analog (AX-B) tool study protein haptenation (AX). AX-B, obtained inclusion biotin moiety at lateral chain AX, showed chemical reactivity identical AX. Covalent modification AX-B was reduced excess AX vice versa, suggesting competition binding same targets. From point view, behaved similarly RAST inhibition studies with sera patients non-selective allergy whereas, expected, poorer AX-selective patients, which recognize chain. Use followed detection allowed observation human serum albumin (HSA) concentrations 100-fold lower that when using detection. Incubation led all previously identified major In addition, some new targets could be detected. Interestingly, intracellular adducts, cell type-specific pattern. This opens possibility following formation fate adducts cells. Thus, may constitute valuable identification high sensitivity well mechanisms involved β-lactams.