College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Global Top5 Program, Ewha Womans University, Seoul 120-750, South Korea.
Department of Chemistry and Nano Science, Ewha Womans University, Seoul 120-750, South Korea.
J Pharm Biomed Anal. 2014 Mar;91:17-23. doi: 10.1016/j.jpba.2013.12.003. Epub 2013 Dec 15.
Human serum albumin (HSA) is the most abundant protein in the human body. HSA injections prepared by fractionating human blood have mainly covered the demand for albumin to treat hypoalbuminemia, the state of low concentration of albumin in blood. HSA in solution may exist in various forms such as monomers, oligomers, polymers, or as mixtures, and its conformational change and/or aggregation may occur easily. Considering these characteristics, there is a great chance of modification and polymer formation during the preparation processes of albumin products, especially injections. The albumin cobalt binding (ACB) test reported by Bar-Or et al. was originally designed to detect ischemia modified albumin (IMA), which contains the modified HSA N-terminal sequence by cleavage of the last two amino acids. In this study, we developed a cobalt albumin binding (CAB) assay to correct the flaws of the ACB test with improving the sensitivity and precision. The newly developed CAB assay easily detects albumin configuration alterations and may be able to be used in developing a quality control method for albumin and its pharmaceutical formulations including albumin injections.
人血清白蛋白（HSA）是人体内最丰富的蛋白质。通过分离人体血液制备的 HSA 注射主要满足了治疗低白蛋白血症（血液中白蛋白浓度低的状态）的白蛋白需求。溶液中的 HSA 可能以单体、寡聚物、聚合物或混合物的形式存在，其构象变化和/或聚集很容易发生。考虑到这些特性，白蛋白产品（尤其是注射剂）的制备过程中很有可能发生修饰和聚合。Bar-Or 等人报道的白蛋白钴结合（ACB）试验最初是为了检测包含通过裂解最后两个氨基酸修饰的 HSA N 末端序列的缺血修饰白蛋白（IMA）而设计的。在这项研究中，我们开发了一种钴白蛋白结合（CAB）测定法，通过提高灵敏度和精密度来纠正 ACB 试验的缺陷。新开发的 CAB 测定法可以轻松检测白蛋白构象的改变，并且可能能够用于开发白蛋白及其药物制剂（包括白蛋白注射剂）的质量控制方法。
