Department of Research and development, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
Department of Research & Development, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Marand, Iran.
Arch Razi Inst. 2024 Feb 1;79(1):102-110. doi: 10.32592/ARI.2024.79.1.102. eCollection 2024 Feb.
Newcastle disease virus (NVD) from the family is a single-stranded negative-sense RNA virus. This infection can affect both domestic poultry and almost all other bird species. It has been considered a very severe difficulty for the poultry industry all over the world. Even though it remains a potential threat to poultry industries, this virus is a powerful oncolytic virus as well. In this study, a process was accomplished to achieve concentrated and highly purified NDV V4 strain particles. Downstream processing of Newcastle virus strain V4 was characterized by amplifying virus in embryonated chicken eggs. Through a sequence of steps, harvesting allantoic fluid, clarification by centrifuge, concentration by ultrafiltration, and size exclusion separation, the reduced volume and pure virus particles were considered for the amount of ovalbumin, hemagglutinin activity, transmission electron microscopy (TEM), electrophoresis, and additionally immunogenicity of prepared antigens. The results presented a high recovery of HA activity in concentrated and purified virus with the removal of ovalbumin and the typical morphology based on TEM. Sepharose CL-4B was determined as the best media among all used resins to purify the virus. Prepared formulations as vaccines demonstrated positive hemagglutinin inhibition for 6 months and stability for 2 years. Strong evidence from organized studies supports the effectiveness of this method in concentrating and purifying intact NDV, which could be valuable in vaccine research, antiserum preparation, or even as an alternative oncotic agent to traditional methods. Despite further studies being conducted, this method can be utilized particularly on a semi-industrial scale to produce various vaccine components.
新城疫病毒（NVD）属于正黏病毒科，是一种单链负义 RNA 病毒。这种感染可以影响家禽和几乎所有其他鸟类。它被认为是全世界家禽业的一个非常严重的难题。尽管它仍然是家禽业的潜在威胁，但这种病毒也是一种强大的溶瘤病毒。在这项研究中，我们完成了一个过程，实现了浓缩和高度纯化的 NDV V4 株颗粒。新城病毒株 V4 的下游处理过程是通过在鸡胚中扩增病毒来实现的。通过一系列步骤，收获羊水，离心澄清，超滤浓缩，和尺寸排阻分离，减少了体积和纯病毒颗粒，考虑到卵清蛋白的量、血凝素活性、透射电子显微镜（TEM）、电泳，以及制备抗原的免疫原性。结果表明，在浓缩和纯化病毒时，HA 活性的回收率很高，同时去除了卵清蛋白，并基于 TEM 呈现出典型的形态。在所有使用的树脂中，Sepharose CL-4B 被确定为纯化病毒的最佳介质。作为疫苗的制剂表现出 6 个月的血凝抑制阳性和 2 年的稳定性。有组织的研究提供了强有力的证据，支持这种方法在浓缩和纯化完整的 NDV 方面的有效性，这在疫苗研究、抗血清制备，甚至作为传统方法的替代溶瘤剂方面都可能具有价值。尽管进一步的研究正在进行，但这种方法可以在半工业规模上用于生产各种疫苗成分。
