Isolation of a somatomedin-binding protein from preterm amniotic fluid. Development of a radioimmunoassay.
Isolation of a somatomedin-binding protein from preterm amniotic fluid. Development of a radioimmunoassay.
复制标题
从早产羊水中分离生长调节素结合蛋白。
DOI：
--
复制
发表时间：
1984
影响因子：
5.8
通讯作者：
H. Guyda
中科院分区：
文献类型：
--
作者：
S. Drop;D. Kortleve;H. Guyda
羊水结合蛋白(AFBP)是一种耐热、耐酸的生长抑素(Sm)结合蛋白，摩尔质量分数为35-40,000，等电点为+/-4.7。它在RRAS中对Sm具有反应性，在Sm生物检测中抑制Sm活性。采用酸-乙醇提取、Sephadex G-150柱层析、高速凝胶过滤层析和圆盘凝胶电泳法从中期人羊水中分离纯化AFBP。通过125I-胰岛素样生长因子II孵育和葡聚糖包被的木炭分离来定量结合活性(每毫克蛋白质的微克当量)。蛋白质回收率不到1%。用纯化的AFBP免疫兔制备AFBP抗血清。用亲和层析法将人血清白蛋白抗体清除。20倍浓缩早产儿房颤和胎儿血清的免疫电泳法显示一条沉淀线。用氯胺-T法标记AFBP。在最终稀释度为1：5000的条件下，AFBP抗血清与加入的~(125)I-AFBP的+/-35%特异性结合。建立了双抗体放射免疫分析方法。30例妊娠中期房颤患者的AFBP水平为148+/-18(扫描电子显微镜)，12例足月房颤患者的AFBP水平为72+/-36mU/ml，胰岛素样生长因子I/Sm-C值(RIA法)均很低(<0.10U/ml)。当血清在pH2.2的Sephadex G-200上层析时，AFBP-RIA活性在一个峰上洗脱，对应的摩尔质量为35-40,000。胎儿血清(胎龄+/-20周)活性最高，成人血清活性最低。AFBP-RIA的建立可能有助于进一步阐明Sm及其结合蛋白在出生前和出生后生长中的生理意义。
Amniotic fluid binding protein (AFBP) is a heat and acid stable somatomedin (Sm)-binding protein with a mol wt of 35-40,000 and an isoelectric point of +/- 4.7. It is reactive in RRAs for Sm and inhibits Sm activity in Sm bioassays. AFBP was purified from midgestational human amniotic fluid (AF) using acid-ethanol extraction, Sephadex G-150 chromatography, high speed gel filtration chromatography, and disc gel-electrophoresis. Specific binding activity (microgram equivalents per mg protein) was quantitated by incubation with 125I-insulin-like growth factor II and dextran-coated charcoal separation. Protein recovery was less than 1%. AFBP antiserum was produced by immunizing rabbits with purified AFBP. The antiserum was cleared of human serum albumin antibodies by affinity chromatography. Immunoelectrophoresis of 20x concentrated preterm AF and fetal serum resulted in one precipitin line. AFBP was labeled by the chloramine-T method. The AFBP antiserum specifically bound +/- 35% of added 125I-AFBP at a final dilution of 1:5000. A double antibody RIA was developed. The AFBP level measured by RIA in midgestation AF (n = 30) was 148 +/- 18 (SEM) and in term AF (n = 12) 72 +/- 36 mu geq/ml. Insulin-like growth factor I/Sm-C values (determined by RIA) in the same samples were uniformly very low (less than 0.10 U/ml). When serum was chromatographed on Sephadex G-200 at pH 2.2, AFBP-RIA activity eluted in one peak corresponding to a mol wt of 35-40,000. Highest activity was found in fetal serum (gestational age +/- 20 weeks) and lowest in serum from adults. The development of the AFBP-RIA may contribute to further elucidation of the physiological importance of Sm and the Sm-binding proteins in pre- and postnatal growth.
垂体功能减退症中的血浆生长调节素结合蛋白：生长激素治疗期间的变化。
DOI：
10.1210/jcem-53-1-100
发表时间：
1981
期刊：
The Journal of clinical endocrinology and metabolism
影响因子：
--
作者：
Hintz,RL;Liu,F;Rosenfeld,RG;Kemp,SF
通讯作者：
Kemp,SF
脐带血浆中生长调节素肽分布和生长调节素结合蛋白含量：与正常和垂体垂体血浆的比较。
DOI：
10.1210/jcem-54-2-223
发表时间：
1982
期刊：
The Journal of clinical endocrinology and metabolism
影响因子：
--
作者：
Borsi,L;Rosenfeld,RG;Liu,F;Hintz,RL
通讯作者：
Hintz,RL