高校化学工程学报
高校化學工程學報
고교화학공정학보
JOURNAL OF CHEMICAL ENGINEERING OF CHINESE UNIVERSITIES
2013年
6期
925-930
,共6页
武强%林东强%翁叶靖%姚善泾
武彊%林東彊%翁葉靖%姚善涇
무강%림동강%옹협정%요선경
双水相系统%混合模式配基%2-巯基-1-甲基咪唑%免疫球蛋白G%分配系数
雙水相繫統%混閤模式配基%2-巰基-1-甲基咪唑%免疫毬蛋白G%分配繫數
쌍수상계통%혼합모식배기%2-구기-1-갑기미서%면역구단백G%분배계수
aqueous two-phase system%mixed-mode ligand%2-mercapto-1-methyl-imidazole%immunoglobulin G%partition coefficient
以2-巯基-1-甲基咪唑(MMI)为混合模式配基，人血免疫球蛋白G(IgG)为模型蛋白，探讨混合模式配基双水相系统分离抗体的新方法，主要包括MMI-聚乙二醇(PEG)的制备和IgG分配研究。首先优化了PEG4000的环氧氯丙烷活化反应条件，环氧氯丙烷和PEG比例为3.5：1，三氟化硼乙醚溶液加量为0.05 mL×g-1 PEG，开环反应时间为8 h；40%氢氧化钠溶液加量为0.15 mL×g-1PEG，闭环反应时间为4 h，环氧基团接入量可达到460μmol×g-1 PEG，偶联配基MMI，得到 MMI-PEG 混合模式配基。进一步考察了 IgG 在 PEG4000/(NH4)2SO4双水相系统中的分配，发现添加 MMI-PEG配基有效提高了IgG分配系数，加入适量NaCl，分配系数达到了17.1。结果表明，混合模式配基双水相萃取可以实现IgG的上相富集，为抗体分离提供了一个新途径。
以2-巰基-1-甲基咪唑(MMI)為混閤模式配基，人血免疫毬蛋白G(IgG)為模型蛋白，探討混閤模式配基雙水相繫統分離抗體的新方法，主要包括MMI-聚乙二醇(PEG)的製備和IgG分配研究。首先優化瞭PEG4000的環氧氯丙烷活化反應條件，環氧氯丙烷和PEG比例為3.5：1，三氟化硼乙醚溶液加量為0.05 mL×g-1 PEG，開環反應時間為8 h；40%氫氧化鈉溶液加量為0.15 mL×g-1PEG，閉環反應時間為4 h，環氧基糰接入量可達到460μmol×g-1 PEG，偶聯配基MMI，得到 MMI-PEG 混閤模式配基。進一步攷察瞭 IgG 在 PEG4000/(NH4)2SO4雙水相繫統中的分配，髮現添加 MMI-PEG配基有效提高瞭IgG分配繫數，加入適量NaCl，分配繫數達到瞭17.1。結果錶明，混閤模式配基雙水相萃取可以實現IgG的上相富集，為抗體分離提供瞭一箇新途徑。
이2-구기-1-갑기미서(MMI)위혼합모식배기，인혈면역구단백G(IgG)위모형단백，탐토혼합모식배기쌍수상계통분리항체적신방법，주요포괄MMI-취을이순(PEG)적제비화IgG분배연구。수선우화료PEG4000적배양록병완활화반응조건，배양록병완화PEG비례위3.5：1，삼불화붕을미용액가량위0.05 mL×g-1 PEG，개배반응시간위8 h；40%경양화납용액가량위0.15 mL×g-1PEG，폐배반응시간위4 h，배양기단접입량가체도460μmol×g-1 PEG，우련배기MMI，득도 MMI-PEG 혼합모식배기。진일보고찰료 IgG 재 PEG4000/(NH4)2SO4쌍수상계통중적분배，발현첨가 MMI-PEG배기유효제고료IgG분배계수，가입괄량NaCl，분배계수체도료17.1。결과표명，혼합모식배기쌍수상췌취가이실현IgG적상상부집，위항체분리제공료일개신도경。
With 2-mercapto-1-methyl-imidazole (MMI) as the mixed-mode ligand and human serum IgG as model protein, new aqueous two-phase system (ATPS) with mixed-mode ligand was developed and used for antibody separation. The preparation of MMI-polyethylene glycol (PEG) and the partitioning behaviors of IgG in ATPS were investigated. Firstly, the activation of PEG was studied, and the optimized conditions were obtained as follow:addition of BF3-C2H5OC2H5 of 0.05 mL×g-1 PEG, ring opening reaction time of 8 h, addition of NaOH (40%(wt)) of 0.15 mL×g-1 PEG and reaction time of 4 h at room temperature. The ratio of ECH and PEG is 3.5:1, and the epoxide content of activated PEG could reach 460μmol×g-1 PEG. Then the PEG-expoxide was coupled with MMI to obtain the MMI-PEG as mixed-mode ligand. Finally, the partition behaviors of IgG in PEG4000/(NH4)2SO4 ATPS were studied. It was found that the MMI-PEG ligand could effectively increase the partition coefficient of IgG in the new ATPS. When some amount of NaCl was added in the system, the partition coefficient could reach 17.1. The results demonstrate that the aqueous two-phase system with mixed-mode ligand could enrich the IgG into the top phase, which provides a new method for antibody separation.
