不同物理和化学因素对重组人角质细胞生长因子2稳定性的影响  

Effects of different physical and chemical factors on stability of recombinant human keratinocyte growth factor-2

作　　者：周鑫[1,2] 王晓杰[1,3,4] 田海山[1,4] 唐禄[1,2] 焦悦[1] 朱家楠[1] 王莘[2] 李校堃[1] 

机构地区：[1]吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林长春130118 [2]吉林农业大学生命科学学院,吉林长春130118 [3]吉林大学公共卫生学院卫生部放射生物学重点实验室,吉林长春130021 [4]吉林省长春市格鲁斯特生物科技有限公司,吉林长春130012

出　　处：《吉林大学学报（医学版）》2010年第3期518-522,共5页Journal of Jilin University：Medicine Edition

基　　金：科技部"重大新药创制"科技重大专项基金资助课题(2008ZX09101-057);长春市应用技术研究计划项目资助课题(2008262);长春市企业科技特派员行动计划项目资助课题(09KT03);长春市科技型中小企业技术创新基金资助课题(09C26212203306)

摘　　要：目的:研究重组人角质细胞生长因子2(rhKGF-2)在不同保存条件下的稳定性,阐明温度、pH值、缓冲溶液及反复冻融因素对rhKGF-2稳定性的影响作用。方法:rhKGF-2在不同温度(-20℃、4℃、25℃、40℃)、pH值(pH4～9)、缓冲溶液(柠檬酸钠-柠檬酸缓冲液、磷酸氢二钠-柠檬酸缓冲液、磷酸盐缓冲液、吉斐氏缓冲溶液)及反复冻融条件下分别保存,采用RP-HPLC法检测rhKGF-2纯度,同时观察外观性状。结果:-20℃、4℃分别保存时,rhKGF-2的各项性质在观察期内都无明显变化;25℃时,rhKGF-2存放28d纯度较0h(100.00%)下降至(68.20±0.14)%;40℃时,rhKGF-2存放9h纯度较0h(100.00%)下降至(4.80±0.39)%。rhKGF-2(pH6.0)在25℃条件下保存90d纯度较0h(100.00%)下降至(79.20±0.12)%,与对照组(0h)比较差异有显著性(P<0.05);pH值为4.0、9.0的rhKGF-2液体在25℃条件下分别保存120h纯度较0h(100.00%)下降至(15.60±0.30)%和(67.30±0.10)%,与对照组(0h)比较差异有显著性(P<0.01)。rhKGF-2在柠檬酸缓冲液中25℃条件下保存60d纯度较0h(100.00%)下降至(79.20±0.15)%,与对照组(0h)比较差异有显著性(P<0.01)。rhKGF-2原液反复冻融后其纯度变化差异无显著性(P>0.05)。结论:rhKGF-2在低温和pH6.0～7.0中性条件下稳定。高温、偏酸性和偏碱性条件下均不利于rhKGF-2稳定。柠檬酸缓冲液有利于rhKGF-2稳定。反复冻融对rhKGF-2稳定性影响不明显。Objective To study the stabilities of recombinant human keratinocytegrowth factor-2（rhKGF-2） deposited at different conditions,and elucidate the influences of temperature,pH,buffer solutions and repeated freeze thawing on rhKGF-2 stability.Methods rhKGF-2 was deposited at different conditions such as temperatures （-20℃,4℃,25℃,40℃）,pH（pH 4-9）,buffer solutions（Citrate buffer solution,Disodium hydrogen phosphate-Citrate buffer,Phosphate buffer,Jifeishi buffer） and repeated freeze thawing,respectively.The purity of rhKGF-2 was determined by RP-HPLC,and the appearance was also observed.Results All properties of rhKGF-2 were not altered during the observation period at -20℃ and 4℃;the purity of rhKGF-2 deposied for 28 d was reduced to （68.20±0.14）% compared with 0 h（100.00%） at 25℃;the purity of rhKGF-2 deposited for 9 h was reduced to （4.80±0.39）% compared with 0 h（100.00%） at 40℃. The purity of rhKGF-2（pH 6.0） deposited for 90 d was reduced to（79.20±0.12）% compared with 0 h（100.00%） at 25℃（P0.05）;the purities of rhKGF-2（pH 4.0,9.0） deposited for 120 h were respectively reduced to （15.60±0.3）% and （67.30±0.1）% compared with 0 h（100.00%）at 25℃（P0.01）.The purity of rhKGF-2（pH6.0） deposited in Citrate buffer solution deposited for 60 d was reduced to （79.20±0.15）% compared with 0 h（100.00%） at 25℃（P0.01）.The purity of rhKGF-2 had no significant change after repeated freeze thawing（P0.05）.Conclusion rhKGF-2 is stable when deposited at low temperature and pH6.0-7.0 neutral conditions.High temperature,acidic and basic conditions are harmful for rhKGF-2 stability.It is profit for rhKGF-2 stability in Citrate buffer solution.There is no impact on rhKGF-2 stability after repeated freeze thawing.

关 键 词：重组人角质细胞生长因子-2 纯度 稳定性 

分 类 号：Q5[生物学—生物化学] Q81