MMP-11多克隆抗体的制备及其法医学意义

目的制备兔抗人基质金属蛋白酶-11(MMP-11)多克隆抗体,建立用MMP-11抗体检测月经血的可行方法,探讨其法医学意义。方法将用基因工程制备的人MMP-11融合蛋白免疫新西兰白兔,饱和硫酸铵法进行抗体纯化。运用蛋白印迹法检测月经血痕、外周血痕、阴道液斑、精液斑、唾液斑和尿液斑,盲测验证该方法的可靠性。结果高效价的兔抗人MMP-11多克隆抗体检测月经血MMP-11蛋白的阳性率为90.48%(93/105),而外周血痕、精液斑、唾液斑、尿液斑和阴道液斑均未检出MMP-11。结论用自制的抗MMP-11多克隆抗体所建立的蛋白印迹法检测月经血中的MMP-11特异性好,灵敏有效,可用于月经血及外周血的鉴别。

Preparation of anti-human MMP-11 polyclonal antibody for forensic application

Objective To obtain the rabbit-anti-human matrix metalloproteinase-11(MMP-11) polyclonal antibody and establish a new immunological method for detecting menstrual blood quickly and accurately in forensic field.Methods The fusion protein,obtained by genetic engineering technique,was used to immunize female rabbits,and antibodies obtained from the immune sera were purified by ammonium sulfate fractionation,their specificity and capability of distinguishing menstrual blood from peripheral blood,vaginal liquid,salivary stain,semen,urine and other biological marks were assayed with Western blot methods.Results The purified specific polyclonal antibodies of rabbit-anti-human MMP-11 were obtained.Tissue specificity of antibodies was evident in menstrual blood by Western blot detection,compared with venous blood,vaginal fluid and other body fluids.Conclusion Western blot analysis with MMP-11 polyclonal antibody can be applied to detect menstrual blood in forensic examination.