噬菌体展示外源多肽的免疫原性研究

Immunological properties of foreign peptides displayed on a Filamentous bacteriophage

利用噬菌体展示技术将丝状噬菌体fd基因 8克隆入 pKK2 2 3-3质粒中 .将白色念珠菌特异表位基因插入到修饰后的质粒载体中 ,并制备了杂合噬菌体 .利用纯化的该表位抗原PA免疫小鼠 ,结果表明该噬菌体 -表位抗原PA在有无佐剂存在的情况下均产生抗体 ,用ELISA方法采用双波长 (4 50 / 6 30nm)检测抗体 ,其在有无佐剂存在的情况下光密度值无明显差别 .免疫后小鼠脾淋巴细胞在有无佐剂的情况下均产生明显增殖 ,未见明显的毒副作用 ,具有良好的安全性 .该方法产生的抗体与传统的合成多肽相比具有方法简便、低价高效等特点 .它将成为生产高效低价疫苗的有效途径 .该噬菌体 -表位抗原PA为真菌疫苗的研制打下基础 .

The geneⅧ of Filamentous bacteriophage was cloned to pKK223-3 vector. A oligonucleotide encoding the specific epitope of Candida albicans was inserted to the above modified plasmid and the hybrid phage was purified. The BALB/c mice was injected with the hybrid phage. It was proved that the Ab against the hybrid phage in the presence or absence of adjuvant indicated no difference. Proliferation assays of T-cells taken from spleen of BALB/C mice injected with the hybrid phage in the presence or absence of adjuvant indicated no difference also. This way of generating Ab against the hybrid phage is simpler and much less expensive than the conventional method of peptide synthesis. It will also be an inexpensive and simple route to the productiion of effective vaccine.