【正文】 male age range 女性年齡范圍為2343年，而男性的年齡范圍 was 30–70 years. 是3070歲。 Enzymatic release of oligosaccharides was car 寡糖的酶法釋放車 ried out by using the deglycosylation kit from BioRad 里德，來自Bio Rad使用deglycosylation套件 Laboratories. 實驗室。 Partially purified SHBG was sub 部分純化的SHBG分 jected to enzymatic oligosacchariderelease prior to SDS jected酶低聚糖釋放前SDS PAGE and Western blotting to determine if any of the PAGE和Western印跡法來確定，如果任何 antibodies recognized either N or Olinked oligosaccha 抗體識別的是N或O型連鎖oligosaccha rides on SHBG. 加載上SHBG。 These experiments were 這些實驗 carried out by using sequential incubations each of 30 min 開展使用每30分鐘連續(xù)孵化 at room temperature with washing steps between each an 在室溫彼此之間的洗滌步驟 tibody incubation. tibody孵化。 Values (nmol/l) are means of dupli 值（nmol / L的）是指dupli cates SD. 蓋茨的SD。 . 。 Nitrocellulose was then washed (three times) 硝化棉是洗（三次） and incubated with antimouse Igperoxidase (1:10 000 in 孵育antimouse免疫球蛋白過氧化物酶（1:10 000 TBS containing % Tween20 and 1% BSA) for 1 h at ％Tween 20與1％BSA）1 h后 20176。C. 白蛋白（BSA）（W / V）20 176。 . 。 This system allowed the purification of up to 10 mg 這個系統(tǒng)允許的凈化至10毫克 of mouse IgG from 200 ml of culture supernatant. 鼠標(biāo)從200毫升培養(yǎng)上清抗體。 C tubes were centrifuged and the supernatant counted. 管，離心，取上清液計算。C. PBS 10分鐘的H DHT在4 176。 A series of exper 一個系列exper， iments measuring SHBG in male and female plasma that iments測量在男性和女性血漿的性激素結(jié)合球蛋白 contained increasing amounts of either added testosterone 或者補充睪酮所載越來越多 (–347 nmol/l) or estradiol (–368 nmol/l) to determine （ nmol / L的）或雌二醇（ nmol / L的），以確定 whether the apparent SHBG level was affected by addition 有無明顯的SHBG水平是受另外 of these steroids were carried out. 這些類固醇進行。 C Plates were then washed four times, 板，然后洗4次， and 100 l monoclonal antibody supernatant 16D5 (1:20 in 和100升的單克隆抗體上清16D5（1:20在 assay buffer) was added to each well for 30 min at 20176。 Plates were then washed and blocked in assay buffer 然后沖板和緩沖液受阻 for 1 h at 20176。 / Steroids 64 (1999) 259–265 /類固醇64（1999）259265 Page 4 第4頁 . 。 Comparison of plasma SHBG levels determined by the Dako ELISA using SHBG polyclonal antibodies and by ligand binding with plasma SHBG Dako公司酶聯(lián)免疫吸附測定血漿SHBG水平，使用性激素結(jié)合球蛋白多克隆抗體和血漿性激素結(jié)合球蛋白結(jié)合配體的比較 levels determined by ELISA using SHBG monoclonal antibodies of either the 11F11/16D5 (panels a and c) or 11F11/2G11 (panels b and d) binations. ELISA法測定的11F11/16D5（面板A和C）或11F11/2G11（板B和D）的組合使用性激素結(jié)合球蛋白單克隆抗體的水平。 The SHBG standard plasma was SHBG標(biāo)準(zhǔn)血漿 derived from pooled, female, third trimester, pregnancy 來自池，女，孕晚期，懷孕 plasma having an SHBG level of 400 nmol/l. 血漿中有400 nmol / L的SHBG水平 The standard 標(biāo)準(zhǔn) plasma was initially diluted 1:1000 in assay buffer and then 等離子最初是在緩沖液稀釋1:1000和然后 serially to generate a series of standards from 0 to 400 連續(xù)生成了一系列的標(biāo)準(zhǔn)，從0到 400 nmol/l. nmol / L的 The plates were again washed, and peroxidase 板塊再次洗凈，過氧化物酶 labeled rabbit antiSHBG serum (1:3000 in assay buffer) 標(biāo)記的兔抗血清性激素結(jié)合球蛋白（緩沖液1:3000） 100 l was added per well for 6 h incubation at 20176。 260 260 JG Lewis et al. JG劉易斯等。 Relative antigenic determinant mapping. 相對的抗原決定簇映射 。 (b) SDSPAGE Western blotting （二）SDS PAGE電泳免疫印跡 of partially purified human SHBG following deglycosylation, lanes 1 and 部分純化的人類SHBG以下deglycosylation，泳道1和 3, probed with antibodies 11F11 and 7H9, respectively. 3，探測與抗體11F11和7H9，分別。 C The plates were then emptied by inversion, and 板倒置，然后清空 Fig. 圖 1. 1。 Briefly, microtitre plates were coated overnight at 4176。 Color development was termi 顏色發(fā)展端子 nated by the addition of 100 l of MH 2 2 SO SO 4 4 per well, 每口井， and absorbance was read at 492 nm. 在492 nm處讀取吸光度。 C The plates were washed four times, and sheep antimouse 板洗四次，羊antimouse Igperoxidase was added (100 l/well at 1:1000 in assay 免疫球蛋白過氧化物酶（100升/以及在1:1000在檢測 buffer) for 1 h at 20176。 C After the plates were emptied, 后板被掏空， 100 l of diluted, pooled, human pregnancy plasma (1:1000 100升稀釋，匯集，人類妊娠血漿（1:1000 in assay buffer) containing elevated levels of SHBG (400 在緩沖液中），其中包含的SHBG水平升高（ 400 nmol/l) was added to each well for an overnight incubation nmol / L的），每孔加入孵育過夜 at 4176。 C過夜100 l/well rabbit antiSHBG serum, diluted in phosphatebuff L /以及兔抗 SHBG血清，稀釋的磷酸 BUFF ered saline (PBS), 29 l of antibody in 10 ml PBS. ERED等效液（PBS），29升的抗體在10毫升PBS。 Screening of supernatants 篩選的上清 ELISA plates (Falcon 3912 microtest III。 PII: S0039128X(98)001196 有價證券投資收益：S0039 128X（98）001196 Page 2 第2頁 . 。 fax: 6433640889. 6433640888傳真：6433640889。 clonal antibodies to human SHBG, both intact and peroxi 對人類SHBG，都完好無損，過氧化物單克隆抗體 daselabeled, were from DAKO Corporation, Carpenteria, 案例分析標(biāo)記，DAKO公司，卡奔塔利亞， CA, USA. 美國加利福尼亞州。 Experimental 實驗 . 。 Here we report the production and char 在這里，我們報告的生產(chǎn)和char acterization of four monoclonal antibodies to human SHBG acterization四個單克隆抗體對人類SHBG that may prove useful tools for the study of SHBG. SHBG研究可能證明有用的工具。性激素結(jié)合球蛋白SHBG，ELISA Plasma sex hormonebinding globulin (SHBG) levels are 血漿性激素結(jié)合球蛋白（SHBG）的水平 widely used together with sex steroid assays to determine 廣泛使用性激素檢測，以確定 the distribution of sex steroid hormones between protein 蛋白質(zhì)之間的分布性類固醇激素 bound and free fractions [1]. 必然和自由的分數(shù)[1] 。 1999愛思唯爾科技公司保留所有權(quán)利。 The assay correlated well with an existing 2day ELISA for SHBG in plasma using與現(xiàn)有的2天ELISA檢測相關(guān)血漿SHBG使用 polyclonal antibodies and also correlated with a dihydrosterone (DHT) ligandbinding assay. 多克隆抗體，并與dihydrosterone（DHT）的配體結(jié)合法相關(guān) 。 Carbohydrate residues do not form part of the antigenic 碳水化合物的殘留物不會形成抗原的一部分 determinants of these two antibodies, although one of these showed increased signal following removal of Nlinked oligosaccharides. 這兩種抗體的決定因素，盡管這些發(fā)現(xiàn)增加的信號，去除N 連接低聚糖。 Immunobiochemistry Laboratory, Clinical Biochemistry Unit, Canterbury Health Laboratories, Christchurch, New Zealand 類固醇及免疫學(xué)生物化學(xué)實驗室，臨床生化組，坎特伯雷衛(wèi)生實驗室，新西蘭基督城 Received 22 May 1998。劉易斯*，彼得A. 愛爾德Steroid amp。 Two of the distinct antibodies were useful for Western blotting and recognized a