A process for preparing an adenovirus-containing preparation for in vivo somatic gene transfer to the myocardium or endothelium of large animals, comprising the steps of
(i) generating adenovirus in a bioreactor under predetermined conditions for obtaining an adenovirus-containing medium;
(ii) optionally separating of adenovirus from the adenovirus-containing medium of step (i) by filtration to obtain an adenovirus-containing filtrate;
(iii) concentrating the adenovirus-containing medium of step (i) or the adenovirus-containing filtrate of step (ii) to obtain a adenovirus-containing concentrate;
(iv) batch purification of the adenovirus-containing concentrate of step (iii) by a Fast Protein Liquid Chromatography (FPLC) in at least two steps at a pressure of less than 0.3 MPa and a volume of at least 200 ml to obtain an adenovirus-containing batch;
(v) optionally concentrating the adenovirus-containing batch of step (iv) to obtain a concentrated adenovirus-containing batch; and
(vi) optionally dialysis of the adenovirus-containing batch of step (iv) or the concentrated adenovirus-containing batch of step (v),
to obtain an adenovirus-containing preparation.
一种用于体内体细胞
基因转移至大型动物心肌或内皮细胞的含腺病毒制剂的制备方法,包括以下步骤
(i) 在
生物反应器中按预定条件产生腺病毒,以获得含腺病毒的
培养基;
(ii) 可选地通过过滤从步骤(i)的含腺病毒
培养基中分离腺病毒,以获得含腺病毒的滤液;
(iii) 浓缩步骤(i)的含腺病毒
培养基或步骤(ii)的含腺病毒滤液,得到含腺病毒的浓缩液;
(iv) 在压力小于 0.3 兆帕、体积至少 200 毫升的条件下,用快速蛋白
液相色谱法 (FPLC)分至少两步对步骤③中的含腺病毒浓缩液进行批量纯化,以获得含腺 病毒的批次;
(v) 可选地浓缩步骤(iv)的含腺病毒批次,以获得浓缩的含腺病毒批次;以及
(vi) 可选地透析步骤(iv)的含腺病毒批次或步骤(v)的浓缩的含腺病毒批次、
得到含腺病毒的制剂。