免疫分析法研究乳輸鐵蛋白

Abstract

本研究主要目的在純化鼠類子宮分泌的Lactotransferrin，並製備專一性高的Anti-Lactotransferrin抗體，作為酵素免疫分析(ELISA)及西方墨點法(Western blotting)方法使用。

酵素免疫分析法的結果知 (l) 血清中，雌雄老鼠都有但雌的略高於雄的，但其量每ug血清中約為1ng的Lactotransferrin。 (2) 生殖器官中，雄老鼠只有貯精囊有且量極高，睾丸、副睾、前攝護腺都沒有，雌老鼠卵巢、陰道、乳線存在有，子宮則未測試到。(3)其他組織腦、肝、心、肺、胰、腎、脾、胸線、十二指腸中，雌雄老鼠二者都存在的有肝、脾、腎，雌的含量略高於雄的含量。另外，雌的胸腺有而雄的沒有。雌的腦中有時可以測到。西方墨點法(Western blotting)的結果，除腦外其餘組織均可測試到有一 65K的蛋白質會和Anti-Lactotransferrin抗體反應。有人報告此65K可能是Lactotransferrin的Low Molecular weight型式，而有待進一步證實。

The study is foused on purification of lactotransferrin from murine uterine fluid and purification of the anti-lactotransferrin antibody. The specific antibody was used for both Enzyme Linked Immunosorbent Assay (ELISA) and Western blotting to measure the distribution of Lactotransferrin among different tissues. Results by ELISA are listed as follows: (1) Lactotransferrin was found in the serum of both female and male mice. The amount of female is about 1ng /ug serum, which is somewhat higher than that of male. (2) In reproductive organs, lactotransferrin can only be found in seminal vescile with high quantity, but not in testis, epididymis and prostate. On the other hand, it could be found in ovary, vagina and mammary gland, but not in uterus of female mice. (3) In other tissues we tested, lactotransferrin was found in liver, spleen and kidney of both male and female mice. Lactotransferrin was present also in thymus of female but not in thymus of male mice. Controversially, data from Western blotting showed the existence of a 65K protein in all of the tested tissues except brain. This 65K protein might be the low molecular weight form of Lactotransferrin .(Teng et al, 1989 Endocrinology 124(2):992-999) The discrepancy of the result of these two methods invites future study.