Study of the allergenic benzypenicilloyl-HSA and its specific separation from human plasma by a pre-designed hybrid imprinted membrane

Benzylpenicilloyl/albumin conjugates (BP-HSA) which have lost all antibacterial activity but possess an immunogenic potential were systematically studied by matrix-assisted laser desorption ionization-time of flight-mass spectrometry, polyacrylamide gel electrophoresis and ultraviolet spectrophotometry. The results showed that the conjugation rate was in the range of 8 : 1-18 : 1. The interaction between human serum albumin (HSA) and penicilloic acid (BPA) was firstly studied by using molecular docking. The results showed that at least 8 activity sites existed on HSA for the conjugation of BPA. BP-HSA was imprinted onto a macroporous chitosan/polyvinylpyrrolidone/carbon nanotubes carrier by using surface precipitation polymerization, and a pre-designed hybrid imprinted membrane (CPC-MIM) was obtained to achieve the specific capturing of BP-HSA from human plasma. The morphologies and physical/chemical properties of membranes were systematically characterized by scanning electron microscopy, atomic force microscopy, Fourier translation infrared spectra, differential scanning calorimeter, thermogravimetric analysis and contact angle measurement. The results indicated that CPC-MIM possessed superior properties for the selective adsorption of BP-HSA and it also had a relatively good thermal stability. The adsorption properties of CPC-MIM were evaluated by comparing with other different constituents of membranes. The results revealed that the maximum adsorptive capacity and imprint factor of CPC-MIM were 0.538 μmol cm −3 and 3.3, respectively. Compared with other proteins, CPC-MIM showed a specific adsorption capacity for BP-HSA, and CPC-MIM could selectively capture BP-HSA from human plasma. This study provides a basis for the further research of the allergic mechanism of penicillins, and the generated hybrid imprinted membrane could potentially be an outstanding separation material for the large-scale continuous selective separation of target proteins from a complex matrix. The proposed principle of preparation of CPC-MIM.