摘要
目的探索合成Aβ1-15重抗原肽疫苗的方法,并对其免疫学活性进行鉴定。方法采用间接偶联法合成8分支Aβ1-15重抗原肽（MAP）,通过高效反相液相色谱（RP-HPLC）、SDS-PAGE以及氨基酸成分分析对其进行初步鉴定。以合成的MAP Aβ1-15C57BL/6小鼠,用ELESA法检测血清特异性抗Aβ抗体。结果间接偶联法合成的MAP Aβ1-15RP-HPLC层析后,呈现为一宽大主峰。SDS-PAGE检测显示,共有8条蛋白带,条带梯度比较均匀,分别为1～8分支的MAP Aβ1-15基酸组份及含量基本与Aβ1-15肽的序列一致。以合成的MAP Aβ1-15免疫C57BL/6小鼠后,可产生高滴度的特异性抗Aβ抗体。结论间接偶联法可成功地合成MAP Aβ1-15,且具有很好的免疫学活性,但合成的MAP产物难以纯化。
AIM： To explore the method of synthesizing the Aβ1-15 multiple antigen peptide （MAP）vaccine and to identify its quality and the immunological activity. METHODS： MAP Aβ1-15 was synthesized by indirect conjugation and analyzed by RP-HPLC, SDA-PAGE and amino acid analysis. Then, C57BL/6 mice were immunized with synthesized MAP Aβ1-15. The specific anti-Aβ antibody in the sera of the immunized mice was identified by ELSA. RESULTS： There was a high and wide peak wave in the RP-HPLC chromatogram. The 8 protein bands identified by SDA-PAGE was identical with 1 to 8 branch of MAP Aβ1-15. The amino acid sequence of synthesized MAP Aβ1-15 was almost similar with the standard. High titer of anti Aβ antibody was obtained in the C57BL/6 mice immunized with MAP Aβ1-15. CONCLUSION： MAP Aβ1-15 could be synthesized successfully by indirect conjugation and the synthesized MAP Aβ1-15 had satisfactory immune activity. But the purification of the synthesized complex remained to be a problem.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2007年第5期391-394,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家重点基础研究发展规划(973)项目(2006cb500700)
国家自然科学基金项目(30400512)
粤港关键领域重点突破项目(20054982210)
广东省自然科学资金(04300218)
广东省社会发展项目(2005B10401047
2006B36004001)
广州市科技计划项目(2005Z3-E4021
2006J1-C0101)
中山市科技局重点攻关项目(4209606)
中国博士后科学基金(2004035603)
