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卵巢抑制素的免疫学研究

Immunological study of ovarian inhibin.

作者信息

Hasegawa Y, Miyamoto K, Fukuda M, Takahashi Y, Igarashi M

出版信息

Endocrinol Jpn. 1986 Oct;33(5):645-54. doi: 10.1507/endocrj1954.33.645.

Abstract

Antisera to purified porcine follicular fluid inhibin of 32 K protein (pFF 32 K inhibin) were raised in rabbits. Increasing doses of an antiserum with high titer could neutralize the maximal suppression of FSH secretion caused by 10 ng of pFF 32 K inhibin from rat anterior pituitary cells in culture. A radioimmunoassay was developed using the antiserum and 125I-labelled pFF 32 K inhibin. Specificity of the antiserum was examined by comparing immunological and biological potencies of various inhibin preparations. Cross-reactivity tests revealed that the antiserum almost recognizes rat ovarian inhibin preparations. The antiserum also recognizes purified bovine follicular fluid inhibin of 32 K protein (bFF 32 K inhibin), but with a cross-reactivity of approximately 20%. Cross-reactivity of human follicular fluid to the antiserum was less than 10%. The antiserum also recognizes inhibin forms of higher molecular weights, 100 K, 80 K, and 55 K proteins, which have previously been identified by gel filtration or SDS-PAGE of crude pFF inhibin preparations under protein-dissociation conditions, indicating that these inhibin forms have common or closely related immunodetermining sites.

摘要

用纯化的猪卵泡液32K蛋白抑制素(pFF 32K抑制素)在兔体内制备抗血清。高滴度抗血清剂量的增加能够中和10 ng培养的大鼠垂体前叶细胞分泌的pFF 32K抑制素对促卵泡激素(FSH)分泌的最大抑制作用。利用该抗血清和125I标记的pFF 32K抑制素建立了放射免疫分析法。通过比较各种抑制素制剂的免疫活性和生物学活性来检测抗血清的特异性。交叉反应试验表明,该抗血清几乎能识别大鼠卵巢抑制素制剂。该抗血清也能识别纯化的牛卵泡液32K蛋白抑制素(bFF 32K抑制素),但其交叉反应率约为20%。人卵泡液与该抗血清的交叉反应率小于10%。该抗血清还能识别分子量更高的抑制素形式,即100K、80K和55K蛋白,这些抑制素形式先前已在蛋白质解离条件下通过粗制pFF抑制素制剂的凝胶过滤或SDS-PAGE鉴定出来,这表明这些抑制素形式具有共同或密切相关的免疫决定位点。

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