Department of Green Technologies, Chemistry & Chemical Engineering Research Center of Iran (CCERCI), Pazhuhesh Blvd., 17th km of Tehran-Karaj highway, 14968-13151 Tehran, Iran; Department of Physico Chemistry, Razi Vaccine & Serum Research Institute (RVSRI), Agricultural Research, Education and Extension Organization (AREEO), P.O. Box 31975/148, Karaj, Iran.
Department of Green Technologies, Chemistry & Chemical Engineering Research Center of Iran (CCERCI), Pazhuhesh Blvd., 17th km of Tehran-Karaj highway, 14968-13151 Tehran, Iran.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Apr 15;1083:222-232. doi: 10.1016/j.jchromb.2018.02.003. Epub 2018 Feb 6.
Vaccine production is a biological process in which variation in time and output is inevitable. Thus, the application of Process Analytical Technologies (PAT) will be important in this regard. Headspace solid - phase microextraction (HS-SPME) coupled with GC-MS can be used as a PAT for process monitoring. This method is suitable to chemical profiling of volatile organic compounds (VOCs) emitted from microorganisms. Tetanus is a lethal disease caused by Clostridium tetani (C. tetani) bacterium and vaccination is an ultimate way to prevent this disease. In this paper, SPME fiber was used for the investigation of VOCs emerging from C. tetani during cultivation. Different types of VOCs such as sulfur-containing compounds were identified and some of them were selected as biomarkers for bioreactor monitoring during vaccine production. In the second step, the portable dynamic air sampling (PDAS) device was used as an interface for sampling VOCs by SPME fibers. The sampling procedure was optimized by face-centered central composite design (FC-CCD). The optimized sampling time and inlet gas flow rates were 10 min and 2 m L s, respectively. PDAS was mounted in exhausted gas line of bioreactor and 42 samples of VOCs were prepared by SPME fibers in 7 days during incubation. Simultaneously, pH and optical density (OD) were evaluated to cultivation process which showed good correlations with the identified VOCs (>80%). This method could be used for VOCs sampling from off-gas of a bioreactor to monitoring of the cultivation process.
疫苗生产是一个生物过程，其中时间和产量的变化是不可避免的。因此，过程分析技术（PAT）的应用将是非常重要的。顶空固相微萃取（HS-SPME）与 GC-MS 联用可以作为过程监测的 PAT。这种方法适用于微生物挥发物（VOCs）的化学特征分析。破伤风是由破伤风梭菌（C. tetani）引起的致命疾病，接种疫苗是预防这种疾病的最终方法。在本文中，SPME 纤维被用于研究培养过程中 C. tetani 产生的 VOCs。鉴定出了不同类型的 VOCs，如含硫化合物，并选择了其中一些作为生物反应器监测疫苗生产过程中的生物标志物。在第二步中，便携式动态空气采样（PDAS）装置被用作 SPME 纤维采样 VOCs 的接口。通过中心复合设计（CCD）对采样程序进行了优化。优化的采样时间和进气流量分别为 10 min 和 2 mL/s。PDAS 安装在生物反应器的排气线上，在孵育期间的 7 天内，通过 SPME 纤维制备了 42 个 VOCs 样品。同时，对 pH 值和光密度（OD）进行了评估，结果表明与鉴定出的 VOCs 有很好的相关性（>80%）。这种方法可以用于从生物反应器废气中采集 VOCs，以监测培养过程。
