Modern genetic techniques, enable the use of animal bioreactor systems for the production and functional enhancement of anti-cancer antibodies. Chicken is the most efficient animal bioreactor for the production of anti-cancer antibodies because of its relatively short generation time, plentiful reproductive capacity, and daily deposition in the egg white. Although several studies have focused on the production of anti-cancer antibodies in egg white, in-depth studies of the biological activity and physiological characteristics of transgenic chicken-derived anti-cancer antibodies have not been fully carried out. Here, we report the production of an anti-cancer monoclonal antibody against the CD20 protein from egg whites of transgenic hens, and validated the bio-functional activity of the protein in B-lymphoma and B-lymphoblast cells. Quantitative analysis showed that deposition of the chickenised CD20 monoclonal antibody (cCD20 mAb) from transgenic chickens increased in successive generations and with increasing transgene copy number. Ultra-performance liquid chromatography (UPLC) tandem mass spectrometry (LC/MS/MS) analysis showed that the cCD20 mAb exhibited 14 N-glycan patterns with high-mannose, afucosylation and terminal galactosylation. The cCD20 mAb did not exhibit significantly improved Fab-binding affinity, but showed markedly enhanced Fc-related functions, including complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) compared to commercial rituximab, a chimeric mAb against CD20. Our results suggest that the transgenic chicken bioreactor is an efficient system for producing anti-cancer therapeutic antibodies with enhanced Fc effector functions.

现代遗传技术使动物生物反应器系统可以用于抗癌抗体的生产和功能增强。鸡是产生抗癌抗体的最有效的动物生物反应器，因为它的产生时间相对较短，繁殖能力强并且每天在蛋清中沉积。尽管一些研究集中于蛋清中抗癌抗体的产生，但是尚未完全进行对转基因鸡衍生抗癌抗体的生物学活性和生理特性的深入研究。在这里，我们报道了从转基因母鸡的蛋清中产生针对CD20蛋白的抗癌单克隆抗体，并验证了该蛋白在B淋巴瘤和B淋巴母细胞中的生物功能活性。定量分析表明，转基因鸡的鸡CD20单克隆抗体（cCD20 mAb）的沉积在连续的世代中都随着转基因拷贝数的增加而增加。超高效液相色谱（UPLC）串联质谱（LC / MS / MS）分析显示，cCD20 mAb表现出14种N-聚糖模式，具有高甘露糖，岩藻糖基化和末端半乳糖基化。与商用利妥昔单抗（一种针对CD20的嵌合mAb）相比，cCD20 mAb并未表现出显着改善的Fab结合亲和力，但显示出显着增强的Fc相关功能，包括补体依赖性细胞毒性（CDC）和抗体依赖性细胞毒性（ADCC）。