摘要
目的 制备兔抗人脂多糖结合蛋白 (LBP)多抗 ,并研究其生物学特性。方法 用LBP基因的真核表达载体pEFBOS LBP作为抗原免疫制备兔抗人LBP的抗血清。用饱和硫酸铵盐析纯化后 ,通过ELISA检测抗体的效价 ;Westernblot鉴定其特异性 ;流式细胞术观察LBP及其抗体对荧光标记脂多糖 (LPS)与单核细胞结合的影响 ;ELISA检测LBP及其抗体对LPS诱导肺泡巨噬细胞分泌TNFα的影响。结果 用LBP基因免疫制备的兔抗LBP多抗与LBP具有良好的结合活性 ;有较高的效价 ;LBP增强LPS与单核细胞结合的作用能被该抗体阻断 ;LBP增强LPS诱导肺泡巨噬细胞分泌TNFα的作用也能被该抗体阻断。
Objective To prepare rabbit anti human lipopolysaccharide binding protein(LBP) polyclonal antibody and study its biological characteristics. Methods The eukaryotic expression vector pEFBOS-LBP was used as antigen to immunize rabbit to prepare antiserum against human LBP. The antiserum was purified by precipitation with ammonium sulphate and its titer tested by ELISA, its specificity identified by Western blot. The effect of LBP and anti LBP antibody on the binding of lipopolysaccharide(LPS) to mononuclear cells was observed by flow cytometry, and the effect of LBP and anti LBP antibody on TNFα secretion of alveolar macrophage induced by LPS was tested by ELISA. Results Rabbit anti human LBP ployclonal antibody prepared with LBP gene was of good binding activity to LBP and of higher titer and could block the effect of LBP. Conclusion Rabbit anti human LBP polyclonal antibody prepared by using LBP gene possesses good biological characteristics.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2002年第11期1307-1309,共3页
Journal of Third Military Medical University
基金
国家自然科学基金重点资助项目 ( 39730 2 10 )