CAJ | 학술논문

应用细胞工程技术研制大菱鲆免疫球蛋白M(immunoglobulin M，IgM)的单克隆抗体并分析其免疫学特性。小鼠骨髓瘤细胞NS0与经IgM免疫的BALB/C小鼠脾细胞融合，经过反复有限稀释法克隆，筛选获得4株抗大菱鲆IgM的单克隆抗体杂交瘤细胞株，分别为B1D1、D5C2、E1B2和F4A1。经小鼠腹水扩大生产后，单抗效价为1∶1.024×106检测灵敏度为32 ng/mL。Western blot分析表明，获得的单抗与IgM重链区特异性结合。交叉结果显示，单抗与大菱鲆血清呈强阳性反应，与褐牙鲆、红鳍东方鲀、许氏平鲉、六线鱼、鲈鱼均呈微弱阳性反应，而与半滑舌鳎、鲤鱼、鲫鱼、草鱼、鳙鱼的血清无交叉反应。本研究制备的大菱鲆IgM单克隆抗体效价高、灵敏度高、特异性强，适合用于大菱鲆免疫学相关研究和生产实际应用。
응용세포공정기술연제대릉평면역구단백M(immunoglobulin M，IgM)적단극륭항체병분석기면역학특성。소서골수류세포NS0여경IgM면역적BALB/C소서비세포융합，경과반복유한희석법극륭，사선획득4주항대릉평IgM적단극륭항체잡교류세포주，분별위B1D1、D5C2、E1B2화F4A1。경소서복수확대생산후，단항효개위1∶1.024×106검측령민도위32 ng/mL。Western blot분석표명，획득적단항여IgM중련구특이성결합。교차결과현시，단항여대릉평혈청정강양성반응，여갈아평、홍기동방돈、허씨평유、륙선어、로어균정미약양성반응，이여반활설탑、리어、즉어、초어、용어적혈청무교차반응。본연구제비적대릉평IgM단극륭항체효개고、령민도고、특이성강，괄합용우대릉평면역학상관연구화생산실제응용。
Four monoclonal antibodies (mAbs) to immunoglobulin M (immunoglobulin M, IgM) of Scophthalmus maximus were produced and characterized. All the mAbs (denominated B1D1,D5C2,E1B2 and F4A1) are showed good anti-turbot IgM reactivity in enzyme linked immunosorbent assay (ELISA) and immunoblotting. Results of ELISA indicated that the titre of the mAb was 1∶1.024 × 106 and the minimum detectable amount of IgM was 32 ng/mL. Results of immunoblotting showed that the mAb reacted with heavy chain of IgM. The mAb showed specificity for Scophthalmus maximus,light specificity for Paralichthys olivaceus,Takifugu ru-bripes,Sebastodes fuscescens,Hexagrammos otakii,Lateolabrax japonicas,and no specifici-ty for Cynoglossus semilaevis,Cyprinus carpio,Carassius cuvieri,Ctenopharyngodon idel-lus,Aristichthys nobilis. The mAbs characterization should prove to be useful for studying the biology of IgM and its practical application.