CAJ | 학술논문

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CD22 mAb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4.5×107 pfu/mL重组病毒，为治疗白血病药物的开发和应用打下了基础。
중조감합항인CD22사개기인공정항체시유일조단태련장량개서원항CD22 mAb적scFv련접기래，재여인IgG1적CH3편단련접소획득중조기인공정항체(cRFB4-CH3)，시목전개발치료B세포계림파류적인원화기인공정항체。위탐토해중조기인공정항체고효표체기술，본연구이용DNA중조기술장함유인IgG1적CH3단적항인CD22사개기인공정항체기인극륭도함신호태적중조간상병독재체pAcSG2중，구건중조질립pAcSG2-cRFB4-CH3병전염도Sf9세포중，구건휴대유중조감합항인CD22사개기인공정항체기인적중조간상병독AcNPV-cRFB4-CH3。통과대해중조독진행PCR화IFA감정，증실획득료가이은정표체항인CD22사개기인공정항체적중조간상병독。이식반시험진일보순화병독，경과3차병독식반극륭，획득독개체도4.5×107 pfu/mL중조병독，위치료백혈병약물적개발화응용타하료기출。
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.