Heldt Caryn L, Gurgel Patrick V, Jaykus Lee-Ann, Carbonell Ruben G
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, North Carolina 27695, USA.
Biotechnol Prog. 2008 May-Jun;24(3):554-60. doi: 10.1021/bp070412c. Epub 2008 May 10.
Virus contamination in human therapeutics is of growing concern as more therapeutic products from animal or human sources come into the market. All biopharmaceutical processes are required to have at least two distinct viral clearance steps to remove viruses. Most of these steps work well for enveloped viruses and large viruses, whether enveloped or not. That leaves a class of small non-enveloped viruses, like parvoviruses and hepatitis A, which are not easily removed by these typical steps. In this study, we report the identification of trimeric peptides that bind specifically to porcine parvovirus (PPV) and their potential use to remove this virus from process solutions. All of the trimeric peptides isolated completely removed all detectable PPV from buffer in the first nine column volumes, corresponding to a clearance of 4.5-5.5 log of infectious virus. When the virus was spiked into a more complex matrix consisting of 7.5% human blood plasma, one of the trimers, WRW, was able to remove all detectable PPV in the first three column volumes, after which human blood plasma began to interfere with the binding of the virus to the peptide resin. These trimer resins removed considerably more virus than weak ion exchange resins. The results of this work indicate that small peptide ligand resins have the potential to be used in virus removal processes where removal of contaminating virus is necessary to ensure product safety.
随着越来越多来自动物或人类源的治疗产品进入市场,人类治疗药物中的病毒污染问题日益受到关注。所有生物制药工艺都要求至少有两个不同的病毒清除步骤来去除病毒。这些步骤中的大多数对包膜病毒和大型病毒(无论是否包膜)都有效。这就留下了一类小型非包膜病毒,如细小病毒和甲型肝炎病毒,这些病毒不容易通过这些典型步骤去除。在本研究中,我们报告了特异性结合猪细小病毒(PPV)的三聚体肽的鉴定及其从工艺溶液中去除该病毒的潜在用途。所有分离出的三聚体肽在前九个柱体积内完全从缓冲液中去除了所有可检测到的PPV,相当于清除了4.5-5.5个对数的感染性病毒。当病毒加入到由7.5%人血浆组成的更复杂基质中时,其中一种三聚体WRW能够在前三个柱体积内去除所有可检测到的PPV,之后人血浆开始干扰病毒与肽树脂的结合。这些三聚体树脂去除的病毒比弱离子交换树脂多得多。这项工作的结果表明,小肽配体树脂有潜力用于病毒去除工艺,在这些工艺中,去除污染病毒对于确保产品安全是必要的。