Frazatti-Gallina Neuza M, Mourão-Fuches Regina M, Paoli Rosana L, Silva Maria L N, Miyaki Cosue, Valentini Elizabeth J G, Raw Isaias, Higashi Hisako G
Laboratório de Raiva, Instituto Butantan, Av. Vital Brasil 1500, 05503-900 S. Paulo, Brasil.
Vaccine. 2004 Dec 9;23(4):511-7. doi: 10.1016/j.vaccine.2004.06.014.
A new rabies vaccine was developed from Vero cells adhered to microcarriers, cultivated in a bioreactor in serum-free medium and infected with the PV/VERO-Paris rabies virus strain. The viral suspensions were concentrated by tangential filtration, purified by chromatography and inactivated with beta-propiolactone. In immunogenicity studies performed in mice immunized with three doses of the new vaccine (seven batches) and the commercial Verorab and HDCV, mean titers of neutralizing antibodies of 10.3-34.6, 6.54 and 9.36 IU/ml were found, respectively. The vaccine presented stability during 14 months at 2-8 degrees C, 30 days at 37 degrees C and 8 h at 45 degrees C. The use of serum-free medium facilitated the downstream process leading to residual cellular DNA values <22.8 pg per dose of vaccine in all produced batches. The effective immunogenicity induced in mice by this vaccine, the degree of purity of the product, the high antigen yield and the reduction of the cost of the product due to the virus production and purification processes, makes this technology very important for countries where rabies presents a great public health problem.
一种新型狂犬病疫苗是由附着在微载体上的Vero细胞制备而成的,这些细胞在生物反应器中于无血清培养基中培养,并感染PV/VERO - Paris狂犬病病毒株。病毒悬液通过切向过滤浓缩,通过层析纯化,并用β-丙内酯灭活。在用三剂新型疫苗(七个批次)以及市售的Verorab和HDCV对小鼠进行免疫的免疫原性研究中,分别发现中和抗体的平均效价为10.3 - 34.6、6.54和9.36 IU/ml。该疫苗在2 - 8℃下可稳定保存14个月,在37℃下可稳定保存30天,在45℃下可稳定保存8小时。使用无血清培养基有利于下游加工,使得所有生产批次的疫苗每剂量残留细胞DNA值<22.8 pg。这种疫苗在小鼠体内诱导的有效免疫原性、产品的纯度、高抗原产量以及由于病毒生产和纯化过程导致的产品成本降低,使得该技术对于狂犬病构成重大公共卫生问题的国家非常重要。