重组疟疾疫苗抗原pfcp2.9发酵 纯化工艺的优化及产业化生产工艺路线的建立-optimization of fermentation and purification process of recombinant malaria vaccine antigen pfcp 2.9 and establishment of industrial production process.docxVIP

上海交通大学生物工程硕士论文 上海交通大学生物工程硕士论文 Shanghai Wanxi ng Bi ophar maceut i cal s Company, consi st i ng of mer ozoi t e Api cal Membr ane Ant i gen- l ( AMA1) domai n III and Mer ozoi t e Sur f ace Pr ot ei n- 1 ( MSP1- 19) , i s bei ng eval uat ed by WHO and PATH/ MVI as a vacci ne candi dat e ant i gen agai nst Pl asmodi um f al ci par um mal ar i a. A Pi chi a past or i s expr essi on syst em cont ai ni ng t he Pf CP2. 9 gene has been const r uct ed. But ant i gen pr ot ei n Pf CP2. 9 i s suscept i bl e t o Pi chi a- pr ot ease degr agat i on. Ther ef or e, by opt i mi zat i on condi t i ons of f er ment at i on and pur i f i cat i on pr ocess, our r esear ch goal s i s t o sol ve t he pr obl em of t he pr ot eol yt i c degr adat i on and r each hi gh- l evel expr essi on and pur i f i cat i on of t ar get ant i gen pr ot ei n. Cr i t i cal pr ocess par amet er s f or f er ment at i on, such as t emper at ur e, pH, met hanol f eed r at e and di ssol ved oxygen et c, wer e eval uat ed and opt i mi zed，and f ound t hat t he f er ment at i on par amet er s ar e pH6. 5, 22℃, EDTANa2 5mmol / L i n t he cul t ur e, t he most amount of Pf CP2. 9 pr ot ei n ar e obt ai ned. The super nat ant i s har vest ed and f ur t her pur i f i ed by mul t i pl e col umn chr omat ogr aphi es, whi ch ar e i n t he or der of hydr ophobi c i nt er act i on, desal t i ng by t angent i al f l ow f i l t r at i on ( TFF) , ani on exchanger , cat i on exchanger and si ze excl usi on, t he pur i t y of Pf CP2. 9 pr ot ei n i s no l ess t han 98%. The f i r st hydr ophobi c i nt er act i on st ep i s t he most i mpor t ant st ep, Not onl y t he t ar get pr ot ei n i s capt ur ed, but al so t he t ar get pr ot ei n i s separ at ed f r om t he Pi chi a- pr ot eases. I n ant i ci pat i on of f ur t her advancement of t hi s candi dat e ant i gen Pf CP2. 9, an opt i mi zat i on and ampl i f i cat