Laboratório de Bioprocessos, Centro de Biotecnologia, Instituto Butantan, Brazil.
Laboratório Especial Piloto de Produtos Biológicos Recombinantes, Instituto Butantan, Brazil.
Carbohydr Polym. 2015 Feb 13;116:67-73. doi: 10.1016/j.carbpol.2014.03.046. Epub 2014 Mar 28.
Haemophilus influenzae type b (Hib) is a human pathogen that causes meningitis in infants worldwide. Capsular polysaccharide linked to a protein has been used as an efficient vaccine, and this approach has reduced the incidence of Hib disease since its inclusion in national immunisation campaigns. The traditional polysaccharide downstream process is based on several ethanol precipitations, treatment with detergents and centrifugation. The aim of this study was to introduce tangential microfiltration (TMF) in the place of centrifugation to simplify handling and to scale up the process. The purity of the polysaccharide was RPNA=1747.2 and RPPrt=196.1 for nucleic acid and protein, respectively, meeting the quality requirements for this polysaccharide. Moreover, the polysaccharide was recognised by at specific antibody, and the ribose and phosphate contents were within the expected limits. Thus, we established a process for the purification of capsular polysaccharide produced by H. influenzae type b that is effective, robust and feasible to be scaling up.
乙型流感嗜血杆菌（Hib）是一种人类病原体，可导致全球婴儿患脑膜炎。与蛋白质相连的荚膜多糖已被用作一种有效的疫苗，自纳入国家免疫接种运动以来，这种方法降低了 Hib 疾病的发病率。传统的多糖下游工艺基于几次乙醇沉淀、用洗涤剂处理和离心。本研究的目的是在不影响多糖质量的前提下，用切向微滤（TMF）代替离心，简化操作并扩大工艺规模。多糖的纯度为 RPNA=1747.2 和 RPPrt=196.1，分别为核酸和蛋白质，符合多糖的质量要求。此外，多糖被特异性抗体识别，核糖和磷酸盐含量也在预期范围内。因此，我们建立了一种有效、稳健且可行的乙型流感嗜血杆菌荚膜多糖的纯化工艺。
