摘要
为了探讨人源抗甲型肝炎(甲肝)病毒scFv-Fc融合抗体在酵母中的表达特性,将获得的人源抗甲肝病毒中和性单链可变区抗体(scFv抗体)基因克隆入含信号肽及人IgG1Fc抗体基因的酵母细胞表达载体中,获得了一株中和性人源抗甲肝病毒pPiscFv-FcHA16融合抗体的分泌表达,并对表达产物进行了纯化。同时对表达产物的生物学特性进行了一系列鉴定。表达的pPiscFv-FcHA16融合抗体为具有不同糖基化形式的同源二聚体,与相应的CHO细胞表达的IgG抗体相比,pPiscFv-FcHA16融合抗体仍保持很好的抗原结合活性,以及与中和性鼠抗甲肝病毒单克隆抗体的竞争抑制能力。同时也保持了对甲肝病毒的体外中和活性。这些结果表明,在酵母中表达的单链可变区(scFv)与IgG1Fc区的融合抗体具有很好的生物学活性,有希望用做体外诊断,用纯化相应的抗原,或者可能用于体内预防与治疗。
The recombinant human neutralizing monoclonal antibody pPiscFv-FcHA16 against hepatitis A virus （HAV） was expressed in Pichia pastoris . The heavy and light chain variable genes of human-derived neutralizing scFv antibody against HAV were cloned into Pichia pastoris expression vector, which already has the signal peptide and human IgG1 Fc fragments fused in it in frame. The pPiscFv-FcHA16 fusion antibody product was purified from the culture medium and well characterized. The expressed pPiscFv-FcHA16 fusion antibody can form a homodimer with different glycosylations. When compared with the same IgG molecule expressed in CHO cells, the expressed pPiscFv-FcHA16 fusion antibody fully retained the specificity of binding to HAV and competition with mouse neutralizing anti-HAV McAb using ELISA, and also retained viral neutralizing activity in vitro . The results of this study demonstrated that the scFv-Fc fusion antibody expressed in Pichia pastoris well retained the biological activities when tested in vitro , pointing to the feasibility of producing scFv-Fc fusions in Pichia pastoris for diagnosis and for purification of antigen in vitro and the possible therapeutic use.
出处
《病毒学报》
CAS
CSCD
北大核心
2005年第5期343-348,共6页
Chinese Journal of Virology
基金
中国比利时合作项目(课题编号:011S0203)
国家高科技项目(课题编号:002AA2Z3339)
