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人α型IFN治疗慢性乙型肝炎、丙型肝炎及白血病有显著疗效。rHuIFN-α2b因其副作用小,疗效高,因而更为广泛使用。其生产过程中亲和层析纯化至关重要。为此我们制备了高特异性单克隆抗体3D9McAb用于分离纯化重组人干扰素α-2b(rHuIFN-α2b),现报道如下。1 材料和方法1.1 rHuIFN-α2b抗原制备 本所自制,用亲和层析柱纯化,比活2.0×10~8IU/mg,SDS-PAGE纯度达99％。1.2 单抗的筛选和制备 取8周龄雌Balb/c小鼠,用适当处理过的抗原免疫5次后进行细胞融合。ELISA法筛选,有限稀释法4次克隆后获得3D9细胞扩大培养,诱生腹水。
Human α-IFN treatment of chronic hepatitis B, hepatitis C and leukemia have a significant effect. rHuIFN-α2b because of its side effects, high efficacy, which is more widely used. Purification by affinity chromatography is crucial during its production. To this end, we prepared a highly specific monoclonal antibody 3D9McAb for the isolation and purification of recombinant human interferon α-2b (rHuIFN-α2b), are reported below. 1 Materials and methods 1.1 rHuIFN-α2b antigen prepared by the home-made, purified by affinity chromatography, specific activity 2.0 × 10 ~ 8IU / mg, SDS-PAGE purity of 99%. 1.2 Screening and preparation of monoclonal antibodies Eight-week-old female Balb / c mice were immunized 5 times with appropriately treated antigens for cell fusion. ELISA screening, limiting dilution method obtained after 4 clones 3D9 cell expansion culture, induced ascites.