The determination of amino acid sequence of human early pregnancy factor and the research on its bioactivity

对人早孕因子(EPF)的生化特性、化学结构和生物活性进行了阐述，特别是EPF在妊娠形成机制中的重要作用。从孕妇尿液中分离纯化了EPF，并测定了其N端氨基酸序列。我们纯化的EPF具有生物活性，具有28KD的分子量和酸性多肽。最近Clarke等人报道了EPF的其他纯化特性，其序列与硫氧还蛋白(Adult型T细胞白血病因子，ADF)的序列一致。Morton等人从人血清中分离纯化的伴侣蛋白10同源肽，其相对分子质量为21kD，酸性多肽与我们分离的多肽相似。为了鉴定这两种EPF的性质，用人工合成的具有EPF部分化学结构的多肽免疫兔，制备了多克隆抗体。对该抗体的生物活性反应(花环抑制活性)和血清学反应(斑点杂交、酶联免疫吸附试验)和生化测定表明，这三种EPF生物活性由完全不同的生化结构组成。

Human early pregnancy factor (EPF) has been elucidated in the biochemical characteristics, chemical structure and the bioactivity specially for the essential role of EPF in the mechanisms of establishment of pregnancy.The purified property from the source of human urine from pregnant women has been determined amino acid sequences from N-terminal of its peptide. Our purified substance with EPF bioactivity had 28KD of molecular weight and acidic peptide.The other purified properties of EPF are from the extract of human placenta had been recently reported by Clarke et al., which had the identified sequences with Thioredoxin (Adult type T cell leukemia factor ; ADF). And Chaperonin 10 homology peptide purified from human serum by Morton et al.which have 21KD of molecular weight and acidic peptide similar with our isolated peptide. In order to identify these rwo EPF properties, the polyclonal antibodies has been made by immunizind synthesized peptides with parts of EPF chemical structure to the rabbit. The response for biological activity (rosette inhibition activity) and serological responses (Dot blot, ELISA), with this antibody, and biochemical determination have demonstrated that these three EPF-bioactive properties were suggested to consists of completely different biochemical structures.