流感病毒裂解工藝的優(yōu)化
發(fā)布時(shí)間：2019-01-29 20:45
【摘要】：目的探討影響流感病毒裂解的因素,優(yōu)化流感病毒裂解工藝。方法分別在不同裂解時(shí)間(Triton X-100裂解1、2、3和4 h)、不同終濃度的裂解劑(終濃度為0.5%、0.6%、0.7%和0.8%的Triton X-100)、不同總蛋白濃度(2 000、3 000和4 000μg/ml)及不同的離子強(qiáng)度[終濃度為0.01、0.05、0.1和0.5 mol/L的PBS(p H 7.2)]下,對(duì)流感病毒進(jìn)行裂解,電鏡下觀察病毒裂解效果,同時(shí)采用免疫單擴(kuò)散法檢測(cè)血凝素含量,Lowry法檢測(cè)總蛋白含量,計(jì)算蛋白含量/血凝素含量比值(裂解后病毒純度)及血凝素回收率,確定最佳裂解工藝參數(shù)。結(jié)果當(dāng)裂解前病毒純化液蛋白含量范圍為2 000~3 000μg/ml,PBS終濃度為0.1 mol/L時(shí),利用終濃度為0.7%~0.8%的Triton X-100裂解流感病毒2 h,可獲得最佳裂解效果。結(jié)論確定了流感病毒最佳裂解參數(shù),優(yōu)化了流感病毒裂解疫苗生產(chǎn)中的裂解工藝。
[Abstract]:Objective to study the factors affecting influenza virus cleavage and optimize the process of influenza virus cleavage. Methods at different pyrolysis time (Triton X-100), the final concentration of Triton X-100 was 0. 5% and 0. 8%, respectively, and the final concentration was 0. 5%, 0. 6% and 0. 8% Triton X-100 at different final concentrations (0. 5%, 0. 6% and 0. 8% Triton X-100), respectively. Different concentrations of total protein (2 000 渭 g/ml and 4 000 渭 g/ml) and different ionic intensities (final concentrations of 0. 01% 0. 05% and 0. 5 mol/L of PBS (p H 7.2) were used to break down the influenza virus. The effect of virus cleavage was observed by electron microscope, hemagglutinin content was detected by immunodiffusion method, total protein content was detected by Lowry method, the ratio of protein content to hemagglutinin content (virus purity after lysis) and the recovery rate of hemagglutinin were calculated. Determine the optimum pyrolysis process parameters. Results when the protein content of purified prelytic virus was in the range of 2 000 渭 g / ml PBS for 0. 1 mol/L, the best lytic effect could be obtained by using 0. 7% Triton X-100 with 0.8% final concentration for 2 h. Conclusion the optimal lysis parameters of influenza virus were determined and the lysis process of influenza virus lytic vaccine was optimized.
【作者單位】： 長春生物制品研究所有限責(zé)任公司疫苗六室;
【基金】：國家科技重大專項(xiàng)(2012ZX09201301-006) 吉林省醫(yī)藥產(chǎn)業(yè)發(fā)展專項(xiàng)資金(YYZX201112)
【分類號(hào)】：R392
[Abstract]:Objective to study the factors affecting influenza virus cleavage and optimize the process of influenza virus cleavage. Methods at different pyrolysis time (Triton X-100), the final concentration of Triton X-100 was 0. 5% and 0. 8%, respectively, and the final concentration was 0. 5%, 0. 6% and 0. 8% Triton X-100 at different final concentrations (0. 5%, 0. 6% and 0. 8% Triton X-100), respectively. Different concentrations of total protein (2 000 渭 g/ml and 4 000 渭 g/ml) and different ionic intensities (final concentrations of 0. 01% 0. 05% and 0. 5 mol/L of PBS (p H 7.2) were used to break down the influenza virus. The effect of virus cleavage was observed by electron microscope, hemagglutinin content was detected by immunodiffusion method, total protein content was detected by Lowry method, the ratio of protein content to hemagglutinin content (virus purity after lysis) and the recovery rate of hemagglutinin were calculated. Determine the optimum pyrolysis process parameters. Results when the protein content of purified prelytic virus was in the range of 2 000 渭 g / ml PBS for 0. 1 mol/L, the best lytic effect could be obtained by using 0. 7% Triton X-100 with 0.8% final concentration for 2 h. Conclusion the optimal lysis parameters of influenza virus were determined and the lysis process of influenza virus lytic vaccine was optimized.
【作者單位】： 長春生物制品研究所有限責(zé)任公司疫苗六室;
【基金】：國家科技重大專項(xiàng)(2012ZX09201301-006) 吉林省醫(yī)藥產(chǎn)業(yè)發(fā)展專項(xiàng)資金(YYZX201112)
【分類號(hào)】：R392
【參考文獻(xiàn)】
相關(guān)期刊論文 前7條
1 王志武 ,盧日峰 ,范洪學(xué);流感病毒的裂解、純化及抗原性研究[J];中國生物制品學(xué)雜志;2002年03期
2 楊U，
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