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利用单克隆抗体对兔睾丸和精子中一种膜抗原的特性鉴定及其抗血清对生育力的影响

Characterization of a membrane antigen from rabbit testis and sperm isolated by using monoclonal antibodies and effect of its antiserum on fertility.

作者信息

Naz R K, Rosenblum B B, Menge A C

出版信息

Proc Natl Acad Sci U S A. 1984 Feb;81(3):857-61. doi: 10.1073/pnas.81.3.857.

DOI:10.1073/pnas.81.3.857
PMID:6366792
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC344937/
Abstract

An antigen was isolated from deoxycholate-solubilized rabbit testis and sperm by using an immunosorbent column containing IgG from a monoclonal antibody (8C10.5) that inhibits fertility. Elution was by stepwise increases in pH (8.0, 10.0, and 11.4), with the pH 11.4 fraction after recycling through the column showing a single band at 63 kilodaltons in slab NaDodSO4/PAGE with a silver stain. The antigen molecule was composed of two subunits, which on two-dimensional PAGE showed many spots within the same molecular size range (50-70 kilodaltons) but differing in charge. The antigen isolated either from testis or sperm showed mainly the same spots. The antigen is periodic acid/Schiff positive and contained 21% carbohydrate. An asialo-derivative of the antigen did not change its characteristics on NaDodSO4/PAGE. This glycoprotein resolved into two types of polypeptides, those binding and those not binding to a lens culinaris lectin column; some of the polypeptides appeared common to both fractions. Murine antiserum against the antigen neither agglutinated nor immobilized rabbit sperm but in immunofluorescence reacted with the plasma membrane of viable rabbit sperm as well as with murine and human sperm. Fertilization of female rabbits inseminated with treated sperm was not affected, but, by 9 days, fertility was significantly reduced (21% of controls). The postfertilization antifertility effect was not due to parthenogenic activation or to polyspermy. The antiserum reacted with one specific band in the one- and two-dimensional gel electrophoretic transfer blot procedure and was unaffected by absorption with different somatic tissues.

摘要

通过使用含有来自抑制生育的单克隆抗体（8C10.5）的IgG的免疫吸附柱，从脱氧胆酸盐增溶的兔睾丸和精子中分离出一种抗原。通过逐步提高pH值（8.0、10.0和11.4）进行洗脱，在通过该柱循环后的pH 11.4级分在平板NaDodSO4/PAGE上经银染显示一条63千道尔顿的单一谱带。抗原分子由两个亚基组成，在二维PAGE上显示在相同分子大小范围（50 - 70千道尔顿）内有许多斑点，但电荷不同。从睾丸或精子中分离出的抗原显示出大致相同的斑点。该抗原对高碘酸/席夫反应呈阳性，含有21%的碳水化合物。该抗原的去唾液酸衍生物在NaDodSO4/PAGE上其特性未改变。这种糖蛋白可分为两种类型的多肽，即与刀豆球蛋白A凝集素柱结合的和不结合的；一些多肽在两个级分中都有出现。针对该抗原的鼠抗血清既不凝集也不固定兔精子，但在免疫荧光中与活兔精子的质膜以及鼠和人的精子发生反应。用处理过的精子对雌兔进行授精，受精不受影响，但到第9天时，生育力显著降低（为对照组的21%）。受精后的抗生育作用不是由于孤雌激活或多精受精。该抗血清在一维和二维凝胶电泳转移印迹法中与一条特异性谱带发生反应，并且不受不同体细胞组织吸收的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/0555b01014d2/pnas00604-0219-h.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/3b1303b9d1f6/pnas00604-0218-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/5fdf18df39a7/pnas00604-0218-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/99dca980d3a0/pnas00604-0219-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/0c0b098a4a2d/pnas00604-0219-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/11afad4b5a5b/pnas00604-0219-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/80a9eb659f9a/pnas00604-0219-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/6784eba1a8b6/pnas00604-0219-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/50fccf50d82b/pnas00604-0219-f.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/38c4f0ec5484/pnas00604-0219-g.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/0555b01014d2/pnas00604-0219-h.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/3b1303b9d1f6/pnas00604-0218-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/5fdf18df39a7/pnas00604-0218-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/99dca980d3a0/pnas00604-0219-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/0c0b098a4a2d/pnas00604-0219-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/11afad4b5a5b/pnas00604-0219-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/80a9eb659f9a/pnas00604-0219-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/6784eba1a8b6/pnas00604-0219-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/50fccf50d82b/pnas00604-0219-f.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/38c4f0ec5484/pnas00604-0219-g.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9531/344937/0555b01014d2/pnas00604-0219-h.jpg

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The serum proteins in multiple myelomatosis.多发性骨髓瘤中的血清蛋白
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Antibody activities of serum and uterine fluid samples from rabbits isoimmunized against sperm fractions.用精子各组分对家兔进行同种免疫后，其血清和子宫液样本的抗体活性。
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