Direct Conjugation of Penicillins and Cephalosporins with Proteins for Receptor Assays of Beta-Lactam Antibiotics

• Published in: Russian journal of bioorganic chemistry 2022-02, Vol.48 (1), p.85-95
• Main Authors: Serchenya, T. S., Harbachova, I. V., Sviridov, O. V.
• Format: Article
• Language: English
• Subjects: Amides; Ampicillin; Antibiotics; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; Cephalosporins; Chemical synthesis; Comparative studies; Conjugates; Conjugation; Life Sciences; Liquid phases; Macromolecules; Mass spectrometry; Monoclonal antibodies; Nanoparticles; Organic Chemistry; Ovalbumin; Penicillin; Proteins; Reagents; Receptors; Review Article; Serum albumin; Solid phases
• ISSN: 1068-1620; 1608-330X
• DOI: 10.1134/S1068162022010125

— Fifteen protein conjugates of penicillins and cephalosporins containing amino- and/or carboxylic groups in the initial structures have been synthesized in the reactions with human serum albumin or ovalbumin using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) or a combination of EDC and N -hydroxysulfosuccinimide at various ratios of the base reagents. A comparative study of conjugates composition and properties has been carried out by UV spectroscopy, mass spectrometry and a ligand-receptor assay. It was shown that the antibiotic residue content of the macromolecules obtained varied from 1 to 22, the beta-lactam cycle remained intact assuring specific interactions of the conjugates with a penicillin-binding protein. In two developed models of receptor bioanalytic systems, an ampicillin conjugate onto a solid phase binds to penicillin-binding protein complexed with a monoclonal antibody, which was detected by an immunoenzyme label in microplate wells or gold nanoparticles on test strips. Conjugated ampicillin binding to the receptor was competitively inhibited by beta-lactam antibiotics added to the liquid phase, and analytical sensitivities relative to penicillin G were 0.05 and 1 ng/mL for microplate and receptor chromatographic systems, respectively.