Cloning and expression of fragments of fibronectin to screen hepatitis b virus surface antigen - binding domains 纖連蛋白的分段表達及與乙肝表面抗原相互作用研究
After dissolving inclusion bodies , renaturing inclusion bodies and further purification by immobilized metal ion affinity chromatography ( imac ) , the fusion protein trx - scfv of electrophoretic purity was obtained , they still possessed antigen - binding activity 經(jīng)變性復性,用固相金屬離子親和色譜法一步分離純化了具有功能的單鏈抗體,在一升的發(fā)酵液中得到78mg的重組蛋白。
Detection of antigen - binding affinity of soluble mg7 scfv elisa was adopted to examine the antigen - binding affinity of soluble mg7 scfv ; competitive elisa was performed to test the inhibitory ratio of soluble mg7 scfv to the binding affinity of mg7 mab with its relevant antigen El舊a拐成惴現(xiàn)2個克隆的可容牲mg7seem明顯的抗原結(jié)合舌隊其a分別為0 832和0 912 ,均高出附性對照( 0
Detection of antigen - binding affinity of mg7 recombinant phage antibody elisa was repeated to confirm the antigen - binding affinity of positive clones screened out in the former procedure ; these positive phages were examined by restriction analysis ( ecor i and hind iii ) ; competitive elisa was performed to test the inhibitory ratio of these positive clones to the binding affinity of mg7mab and its relevant antigen , the positive dones possessing apparent inhibitory effect were singled out for later use X陽性克到駛知烘扳原kbbjg )濁對陽性克隆進行限制隴臥賜析( uwi和hedlll )鑒定三用競爭elisatoljmg7重組噬菌體抗體性克隆對mg7單扶與其相應(yīng)抗原結(jié)合忙的喇率,從中j ) ed出對mg7單抗與期??乖Y(jié)合有抑余j效應(yīng)的克隆用于進一涉研究。
( 4 ) in artificial pepsin digestive system with ph2 . 0 , due to the acid denaturation , the pigg was hydrolyzed wholly , in the system with ph3 . 0 and 4 . 0 , although not all entirely pigg could be detected , but the pigg antibody activities against e . coli still existed without changing , this indicates that the f ( ab " ) 2 with antigen - binding activity still remain complete during digesting process ( 4 )在ph2 . 0的人工胃液消化體系中,由于酸變性pigg全部水解;在ph3 . 0和ph4 . 0的人工胃液消化體系中,盡管檢測不到所有的pigg ,但pigg仍可與e . coli特異性結(jié)合且凝集價未發(fā)生變化,表明在消化過程中具有抗原結(jié)合活性的f ( ab ) _ 2片段仍可完整地保留下來。