摘要
目的:以卵清蛋白(OVA)为模型抗原蛋白,考察其体外棕榈酸化修饰对免疫反应的影响。方法:根据OVA与棕榈酸N-羟基琥珀酰亚胺酯的不同摩尔比进行投料反应,同时比较不同pH条件对反应产物的影响,并采用SDSPAGE、圆二色谱和表面疏水值测定等技术和方法进行表征;修饰产物再经体外释放分析和体内免疫研究,比较和判断OVA的棕榈酸修饰及其接种方式(皮下或肌肉注射)对免疫动物血清中特异性抗体产生及滴度水平的影响。结果:OVA在pH6的磷酸缓冲液中进行棕榈酸修饰所得产物的表面疏水值最高,且二级结构无明显改变,可提高未修饰OVA免疫动物血清特异性抗体的浓度,与接种方式无关。结论:体外棕榈酸修饰可以改变抗原蛋白的疏水性,增强免疫效果,有助于减少疫苗抗原和佐剂的使用量。
Objective:Ovalbumin(OVA)was used as a model antigen to investigate the effect of palmitoylation on immunogenicity.Methods:The reactions were performed based on different molar ratios between OVA and palmitate N-hydroxysuccinimide ester under the different pH conditions.SDS-PAGE,circular dichroism(CD)and surface hydrophobicity were applied for the characterization of modified products.The palmitoylated OVA was subjected to in vitro release assay and in vivo immunological studies to examine the effect of palmitoylation and the vaccination strategies(subcutaneous and intramuscular)on the specific antibody in the serum of immunized animals.Results:Palmitoylated OVA produced in phosphate buffer pH at 6 gives the highest surface hydrophobicity and the modification causes no significant change in secondary structure from CD spectra.Palmitoylation could increase the concentration of specific antibodies in the serum of immunized mice regardless of the inoculation method.Conclusion:Palmitoylation can modify the hydrophobicity of the antigenic protein,resulting in enhanced immunogenicity and reduced dosage of antigens and adjuvants.
作者
张义浜
陈智峰
马晓英
ZHANG Yi-Bang;CHEN Zhi-Feng;MA Xiao-Ying(School of Pharmacy,Jiangsu University,Zhenjiang 212013,China)
出处
《生物技术通讯》
CAS
2020年第5期537-541,共5页
Letters in Biotechnology
关键词
卵清蛋白
棕榈酸化
蛋白修饰
表面疏水值
免疫原性
ovalbumin
palmitoylation
protein modification
surface hydrophobicity
immunogenicity
