Herr J C, Summers T A, McGee R S, Sutherland W M, Sigman M, Evans R J
Biol Reprod. 1986 Oct;35(3):773-84. doi: 10.1095/biolreprod35.3.773.
A novel sperm-coating antigen from the human seminal vesicles was discovered. We identified a monoclonal antibody MHS-5, recognizing an epitope with characteristics of a forensic semen marker: conservation in all vasectomized or normal semen samples tested (421); absence in all human tissues or biological fluids other than semen; and immunolocalization on the surface of ejaculated sperm. Western blots of ejaculates allowed to liquefy for 5 min demonstrated the MHS-5 epitope to be located on peptides of a wide range of molecular masses from 69 to 8 kDa. After 15 h of semen liquefaction, immunoreaction peptides of higher molecular mass were undetectable in semen, while peptides of lower molecular mass from 8 to 21 kDa retained antigenicity. Three peptides of 10, 11.9, and 13.7 kDa were the most immunoreactive species in semen liquified for 15 h. Using the MHS-5 monoclonal, an enzyme-linked immunosorbent assay (ELISA) was developed sensitive to 1 ng of seminal protein. This assay showed that the MHS-5 antigen was undetectable in semen of common domestic animals and monkeys but was present in chimpanzee, gorilla, and orangutan semen. ELISA of homogenates from human organs and reproductive tissues demonstrated the antigen only in samples of seminal vesicles. Epididymal sperm obtained at vasovasostomy lacked the MHS-5 epitope, a fact that, together with immunolocalization on ejaculated sperm, demonstrated that the MHS-5 antigen functions as a "sperm-coating antigen." The MHS-5 monoclonal detected semen in sexual-assault evidence obtained six months previously and in mixtures of semen with vaginal or cervical fluid. Assay systems employing the MHS-5 monoclonal may be useful for identification of semen in sexual-assault casework. The MHS-5 epitope resides on novel seminal vesicle-specific peptides whose functions, aside from sperm coating, are uncharacterized.
发现了一种来自人类精囊的新型精子包被抗原。我们鉴定出一种单克隆抗体MHS - 5，它识别一种具有法医精液标志物特征的表位：在所检测的所有输精管结扎或正常精液样本中均存在（421份）；在精液以外的所有人体组织或生物体液中均不存在；且在射出精子表面有免疫定位。对液化5分钟的射精进行的蛋白质印迹分析表明，MHS - 5表位位于分子量范围从69至8 kDa的多种肽段上。精液液化15小时后，精液中无法检测到高分子量的免疫反应性肽段，而分子量从8至21 kDa的低分子量肽段仍保留抗原性。10 kDa、11.9 kDa和13.7 kDa的三种肽段是液化15小时的精液中免疫反应性最强的物质。使用MHS - 5单克隆抗体开发了一种对1 ng精液蛋白敏感的酶联免疫吸附测定（ELISA）。该测定表明，普通家畜和猴子的精液中未检测到MHS - 5抗原，但黑猩猩、大猩猩和猩猩的精液中存在该抗原。对人体器官和生殖组织匀浆进行的ELISA表明，该抗原仅存在于精囊样本中。输精管吻合术时获取的附睾精子缺乏MHS - 5表位，这一事实与射出精子上的免疫定位一起，证明MHS - 5抗原作为一种“精子包被抗原”发挥作用。MHS - 5单克隆抗体在六个月前获得的性侵犯证据以及精液与阴道或宫颈液的混合物中检测到了精液。采用MHS - 5单克隆抗体的检测系统可能有助于性侵犯案件中精液的鉴定。MHS - 5表位位于新型精囊特异性肽段上，除了精子包被功能外，其功能尚未明确。
