Plasma fractionation industry is by far the largest protein pharmaceutical provider, but there are still some plasma components in its industrial waste liquid that have not been utilized. This study aimed to develop a simple and efficient method for plasma protein recovery from Cohn fraction V supernatant (FVS), an effluent containing about 40% ethanol. A new affinity chromatography medium was synthesized with a fatty acid ligand. When the medium was applied to recovery of human serum albumin (HSA) from FVS at physiological pH7.4, the process was unsuccessful due to substantial decrease in capacity in the presence of high ethanol concentration. Nevertheless, change of pH from 7.4 to 4.2 emerged an improved adsorption capacity. The carboxyl group of the ligand began to act as cationic ion exchange role. Both HSA and α2HS-glycoprotein were adsorbed by the column, but α2HS-glycoprotein could be eluted by increasing pH from 4.2 to 7.4, while HSA was retained by the column and could only be eluted by addition of fatty acid. Therefore, the adsorption of albumin under pH 4.2 is charge-induced affinity adsorption, not simple ion exchange. The so-called dual-mode adsorption depends not only on the chromatographic medium but also on the separated object and environment. HPSEC showed that the purity of recovered HSA was greater than 98%. Circular dichroism and fluorescence spectra were consistent with that of the commercial product. Furthermore, the measurement by isothermal titration calorimetry showed that the separated HSA still maintained the binding activities with the ligands of warfarin and naproxen. It is therefore possible to directly recover high-purity and high-quality human serum albumin from the effluent of plasma fractionation industry by one-step chromatography.

血浆分馏行业是迄今为止最大的蛋白质制药供应商，但其工业废液中仍有一些血浆成分未得到利用。本研究旨在开发一种简单有效的方法，用于从 Cohn 组分 V 上清液 (FVS) 中回收血浆蛋白，FVS 是一种含有约 40% 乙醇的流出物。用脂肪酸配体合成了一种新的亲和层析介质。当将该培养基用于在生理pH7.4下从FVS中回收人血清白蛋白(HSA)时，由于在高浓度乙醇存在下容量大幅下降，该过程失败。然而，pH 从 7.4 变为 4.2 时，吸附能力有所提高。配体的羧基开始充当阳离子离子交换作用。 HSA和α2HS-糖蛋白均被柱吸附，但α2HS-糖蛋白可以通过将pH从4.2增加到7.4来洗脱，而HSA被柱保留并且只能通过添加脂肪酸来洗脱。因此，pH 4.2下白蛋白的吸附是电荷诱导的亲和吸附，而不是简单的离子交换。所谓双模式吸附不仅取决于色谱介质，还取决于分离的对象和环境。 HPSEC显示回收的HSA纯度大于98%。圆二色性和荧光光谱与商业产品一致。此外，等温滴定量热法测量表明，分离的HSA仍保持与华法林和萘普生配体的结合活性。因此可以通过一步层析从血浆分离行业的废水中直接回收高纯度、高质量的人血清白蛋白。