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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。
In order to investigate the expression characteristics of human anti-Hepatitis A (Hepatitis A) virus scFv-Fc fusion protein in yeast, the obtained human anti-Hepatitis A virus neutralizing single chain variable region antibody (scFv antibody) Peptide and human IgG1 Fc antibody gene yeast cell expression vector obtained a neutralizing human anti-hepatitis A virus pPiscFv-FcHA16 fusion antibody secretion expression, and the expression product was purified. At the same time, a series of identification of the biological characteristics of the expressed product was carried out. The expressed pPiscFv-FcHA16 fusion antibody was a homodimer with a different glycosylation pattern and the pPiscFv-FcHA16 fusion antibody still retained good antigen binding activity as compared to the corresponding CHO antibody expressed by CHO cells, And competitive murine anti-hepatitis A virus monoclonal antibody competitive inhibition. At the same time, the in vitro neutralizing activity of hepatitis A virus is also maintained. These results indicate that the fusion antibodies of the single chain variable region (scFv) expressed in yeast and the IgG1 Fc region have good biological activity and are expected to be used for in vitro diagnosis, purification of the corresponding antigen, or possibly for in vivo prevention With treatment.