与海洛因有完全交叉反应的抗吗啡单抗的制备  

Preparation of anti-morphine monoclonal antibodies with complete cross reactivity with heroin

作　　者：李琳[1] 刘洁[2] 朱平[2] 徐江平[1] 富宁[2] 

机构地区：[1]南方医科大学药理学教研室,广东广州510515 [2]南方医科大学免疫学教研室,广东广州510515

出　　处：《第一军医大学学报》2005年第7期833-836,共4页Journal of First Military Medical University

基　　金：广州市科技攻关项目(2002Z3-E4102)~~

摘　　要：目的制备吗啡的单克隆抗体(mAb)并鉴定抗体的特性。方法6-琥珀酰化吗啡与Bluecarrier(BC)载体蛋白交联制备6-琥珀酰化吗啡酯(M-6-S-BC),用其免疫BALB/c小鼠,取其脾细胞与Sp2/0骨髓瘤细胞融合制备杂交瘤,经筛选和3次克隆化,从制备的腹水中纯化单克隆抗体。结果获得4株杂交瘤细胞G1、G3、H4和B1,其分泌单抗的亚型均为IgG1型。经ELISA鉴定,该单克隆抗体可结合游离的吗啡、海洛因及可待因,而且对海洛因的结合优于吗啡,该单抗不与非阿片类药物结合。结论所制备的吗啡单克隆抗体可特异性结合吗啡等阿片类药,将为建立体外吗啡及海洛因的非同位素检测方法奠定基础。Objective To prepare and characterize anti-morphine monoclonal antibodies (mAbs) with complete cross reactivity with heroin.Methods Morphine-6-succinyl was conjugated with Blue carrier (BC) in the presence of carbodi imide.BALB/c mice were immunized with the holoantigens (M-6-S-BC) and Freund adjuvant,and hybridoma was generated with mouse spleen cells and Sp2/0 myeloma cells.After 3 fusions and cloning,4 hybridoma cell lines secreting monoclonal antibodies to morphine were obtained and the mABs purified from the mouse ascites followed by characterization with enzyme-linked immunosorbent assay (ELISA).Results Four hybridoma cell lines secreting anti-morphine mAbs were established and designated as G1,G3,H4 and B1,respectively.All the mAbs belonged to IgG1 isotype and could bind to heroin better than to morphine and codeine,which were identified by competitive inhibition ELISA.Conclusion The anti-morphine mAbs can bind with the opioids,especially heroin,and can be useful for sensitive detection of opioids.

关 键 词：吗啡 单克降抗体 竞争性抑制 

分 类 号：R392[医药卫生—免疫学]