建立一种快速、简单、高效的从含有多种细胞类型的生物样本（混合斑）中鉴定和分离精子的方法是法医学急需解决的难题之一。本项目是基于精子在附睾中的成熟需要持续的表面修饰为理论基础，针对精细胞膜表面或膜内的特异蛋白，利用抗原-抗体反应分离混合斑中的精细胞。人类载脂蛋白6（hLCN6）是一种附睾分泌蛋白，它能结合到精子的头部和尾部，与精子成熟有关。我们从分泌抗hLCN6单克隆抗体的杂交瘤细胞中克隆抗体可变区基因，装入表达载体后，在宿主细胞中表达并纯化重组抗体。根据抗体所带标签不同,采用亲和层析或流式细胞术的方法方便快捷的将混合生物样本中的精细胞分离纯化出来。分离的精子可用于体外检测或直接裂解用于提取男性DNA并进行遗传分型。本研究可解决目前精子分离中的特异性差和体细胞污染等问题，不仅为性侵案件中法医学鉴别并分离精细胞及后续的个体识别提供新方法，而且为依据抗原-抗体反应分离混合斑中的精细胞开辟新途径。

To establish a rapid, simple and efficient method for the identification and isolation of sperm cells from biological samples containing multiple cell types (mixed stain）is an urgent and difficult task in forensic science. This project is based on the theoretical basis that sperm maturation in epididymis requires successive surface modifications, aimed at the specific proteins located on the cell surface or within the sperm cell and utilizing immuno-binding reaction to separate sperm cells from mixed stain samples. Human lipocalin 6 (hLCN6) is an epididymal secreted protein, capable of binding to the head and tail of spermatozoa and associated with sperm maturation. We plan to clone the variable region genes of anti-hLCN6 monoclonal antibody from hybridoma cells. After connecting the genes into the expression vector, we express and purify the recombinant antibody against hLCN6 from the host cells. According to the tag carried by different recombinant antibodies, sperm cells from mixed biological samples can be isolated and purified in a convenient and efficient way by using affinity chromatography or flow cytometry method. Isolated sperm cells can be used for in vitro analysis or directly lysed for the extraction of male specific DNA and genotyping. The study resolves the problems of low specificity and somatic cell contamination existed in the current methods of spermatozoa isolation, which not only provides a new method for the forensic identification and isolation of sperm cells for subsequent individual identification in sexual assault cases, but also opens up new research avenues for the separation of sperm cells from mixed stains based on immuno-binding reaction.