查看更多>>摘要：As a general approach Fab immobilized immunoaffinity cryogels for human serum albumin purification is presented in this article. The directed immobilization of antibodies, which have been used as diagnostic and therapeutic agents in applications recently, has brought a different perspective to the purification method. In our study to shed light on this method, anti-HSA Fab fragment immobilized poly(hydroxylethyl methacrylate-N-methacryloyl-(l)-cysteine) (PHEMAC-Fab) and PHEMAC cryogels were prepared for the purification of human serum albumin (HSA). The specific surface areas of the PHEMAC-Fab and PHEMAC cryogels were calculated as 38.6 m(2)/g, and 20.0 m(2)/g, respectively. In comparison, the maximum swelling ratio was observed in the PHEMAC-Fab cryogel and more water was absorbed than the PHEMAC cryogel. While the macropore ratio in the PHEMAC-Fab cryogel is 54.3%, the macropore ratio of the PHEMAC cryogels is 41.6%. The results obtained here showed that anti-HSA cryogel prepared by immobilization of Fab fragments can be an alternative to traditional immunoaffinity techniques for HSA purification.
查看更多>>摘要：Metal-organic frameworks (MOFs), defined as a class of microporous hybrid materials, are established by coordination of metal cations with functional organic ligands. In addition to outstanding porosity and large surface area, the structures and functions of MOFs can be designed and adjusted based on different destinations, which would be employed as stationary phases in various chromatographic modes. Pepsin, a class of protein, has been investigated as chiral selector owing to their unique interactions with analytes based on chiral or affinity selectivity. In this work, MOF-5 was exploited to grow on the support of poly (glycidyl methacrylate)-co(ethylene dimethacrylate) [poly(GMA-co-EDMA)] monoliths by layer-by-layer self-assembly method. Pepsin was subsequently bonded with the carboxyl group of MOF-5 through amidation reaction. The ultimate pepsin@MOF5@poly(GMA-co-EDMA) column was applied for enantioseparation of six basic chiral drugs by capillary electrochromatography with good resolution and repeatability. Compared with the pepsin modified monolithic column, the newly prepared column with MOF-5 shows significantly enhanced enantiomeric resolution, which reveals that MOFs-modified capillary monolithic columns lead a promising road to separation of racemates by chromatography.
查看更多>>摘要：Therapeutic monoclonal antibodies (mAbs) represent a very important class of the current biopharmaceutics. The great complexity of their structure made necessary the use of different analytical approaches for assessing different physico-chemical properties. In this work, weak cation exchange (WCX) high performance liquid chromatography with diode array detection ((WCX)HPLC/DAD) is used to assess the charge variant profile. The method here developed combined the effect of ionic strength and controlled pH gradient and allows for the charge variants analysis of the five mAbs studied, namely bevacizumab (BVZ), cetuximab (CTX) infliximab (INF), rituximab (RTX) and trastuzumab (TTZ), which are among the most used mAbs worldwide. The differences in the charge variants in the natural isoforms of the mAbs promoted characteristic WCX chromatograms for each of mAbs that can be also useful for identification purposes. These chromatograms have provided to be suitable for tracking changes in the charge variants of each mAb analyzed both in controlled degraded and in stabilities study along time of in-use samples solutions at 2 mg/mL in 0.9% NaCl stored refrigerated (at 4 ?C) and frozen (-20 ?C) for two months. The results obtained indicated different stabilities of these mAbs, all IgG1, against degradation by different stressed environmental conditions and in-use stability along two months.
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Elsevier