人溶菌酶基因免疫抗体的制备及其初步应用  

PREPARATION OF ANTIBODY TO HUMAN LYSOZYME BY GENE IMMUNIZATION AND ITS PRIMARY APPLICATION

作　　者：李国才[1] 居杏珠[1] 黄永生[1] 孙怀昌[2] 张泉[2] 朱秋菊[2] 颜新敏[2] 

机构地区：[1]扬州大学医学院,江苏扬州225001 [2]扬州大学兽医学院,江苏扬州225009

出　　处：《实用临床医药杂志》2005年第7期33-34,45,共3页Journal of Clinical Medicine in Practice

基　　金：国家教育部骨干教师培养计划(2000);江苏省高新技术项目(BJ2001315)

摘　　要：目的制备人溶菌酶(hLYZ)基因免疫抗体。方法用多聚乙酰亚胺包裹重组质粒pcDNA3LYZ,经肌肉注射途径免疫BALB/c小鼠,以间接ELISA检测免疫血清中hLYZ抗体的效价,所得抗体用Western-blotting法检测转基因小鼠乳汁中重组hLYZ的表达情况。结果2次免疫后,小鼠体内产生了hLYZ特异性抗体,效价达1∶800,用此抗体为第一抗体在转基因小鼠乳汁中检测到分子量与正常hLYZ相同的重组hLYZ。结论用基因免疫方法在小鼠体内制备了可用于检测的hLYZ抗体。Objective To prepare antibody against human lysozyme（hLYZ） by gene immunization methods. Methods The recombinant expression vector pcDNA3LYZ was injected into BALB/c mice after mixed with 25 kDa polyethylenimine, and the obtained serum was detected by ELISA. The antiserum was used to assay the expression of recombinant hLYZ in milk of hLYZ transgenic mice by Western-blotting. Results After immunized twice, the BALB/c mice produced antibody of 1 ： 800 against hLYZ. By the prepared antiserum as the first antibody, the recombinant hLYZ expressed in milk of the transgenic mice was turned to have the same molecular weight of 14.7 kDa as the normal hLYZ. Conclusion Antibody against hLYZ that could be used as detection reagent was prepared successfully.

关 键 词：人溶菌酶 基因免疫 抗体 

分 类 号：R446[医药卫生—诊断学]