重组嵌合抗人CD22四价基因工程抗体在昆虫杆状病毒中的表达  

Expression of Recombinant, Chimeric and Genetically Engineered Tetravalent Anti-human CD22 Antibody in Baculovirus Expression System

作　　者：马玲[1] 邵帅[1] 潘汉世[1] 陈凤莲[1] 黄红梅[1] 石胜[1] 吴健敏[1] 

机构地区：[1]广西兽医研究所,广西畜禽疫苗新技术重点实验室,南宁530001

出　　处：《基因组学与应用生物学》2013年第3期314-318,共5页Genomics and Applied Biology

基　　金：广西"新世纪十百千人才工程"人选专项经费(桂人办发〔2005〕32号);广西水产畜牧兽医局项目(桂鱼牧科2003-65-06);广西自然科学基金(桂科自0991220)共同资助

摘　　要：重组嵌合抗人CD22四价基因工程抗体是由一条短肽链将两个鼠源抗CD22mAb的scFv连接起来,再与人IgG1的CH3片段连接所获得重组基因工程抗体(cRFB4-CH3),是目前开发治疗B细胞系淋巴瘤的人源化基因工程抗体。为探讨该重组基因工程抗体高效表达技术,本研究利用DNA重组技术将含有人IgG1的CH3段的抗人CD22四价基因工程抗体基因克隆到含信号肽的重组杆状病毒载体pAcSG2中,构建重组质粒pAcSG2-cRFB4-CH3并转染到Sf9细胞中,构建携带有重组嵌合抗人CD22四价基因工程抗体基因的重组杆状病毒AcNPV-cRFB4-CH3。通过对该重组毒进行PCR和IFA鉴定,证实获得了可以稳定表达抗人CD22四价基因工程抗体的重组杆状病毒。以蚀斑试验进一步纯化病毒,经过3次病毒蚀斑克隆,获得毒价达到4.5×107pfu/mL重组病毒,为治疗白血病药物的开发和应用打下了基础。cRFB4-CH3 is a recombinant, chimeric and genetically engineered tetravalent anti-human CD22 anti-body, obtained by linking two scFvs of CD22 McAb with a short peptide chain, and then linked with the CH3 of human IgG1. At present, it was developed to cure B-cell lymphoma. In order to investigate the high expression of it, the gene of cRFB4-CH3 was cloned into baculovirus expression vector pAcSG2 so as to construct recombinant plasmid pAcSG2-cRFB4-CH3, which was then transfected into Sf9 cells to obtain recombinant baculovirus AcN-PV-cRFB4-CH3. Identification by PCR and IFA suggested that AcNPV-cRFB4-CH3 could express cRFB4-CH3 steadily. And after 3 purification cycles by plaques assay, the titer of the purified AcNPV-cRFB4-CH3 reached 4.5í107 pfu/mL. The results of this research will provide a basis for the development and application of therapy of B-cell lymphoma.

关 键 词：CD22 人源化 基因工程抗体 杆状病毒 表达 

分 类 号：Q789[生物学—分子生物学]