Iwamori M, Hirota K, Utsuki T, Momoeda K, Ono K, Tsuchida Y, Okumura K, Hanaoka K
Department of Biochemistry, The University of Tokyo, 7-3-1 Hongo, Tokyo, Bunkyo-ku, 113.
Anal Biochem. 1996 Jun 15;238(1):29-33. doi: 10.1006/abio.1996.0245.
By TLC-immunostaining with the monoclonal antibody VJ-41, which preferentially reacted with sphingomyelin (Sm) and disaturated fatty acid-containing phosphatidyl choline (DSPC), Sm and surfactant phospholipid dipalmitoyl PC were only detected in the lipid extracts from human amniotic fluid. The method was useful in the selective and simultaneous determination of surfactant phospholipid and Sm concentrations in the amniotic fluids to determine the level of maturity of the lungs of the fetus. By measuring the density of spots visualized by TLC-immunostaining, we detected Sm at a sensitivity two times higher than that for dipalmitoyl PC using the antibody. More than 50 ng of dipalmitoyl PC and Sm was detected on the same TLC plate and the standard curves were linear up to 1 microgram of phospholipids. The method was applied to determine the surfactant phospholipid/Sm ratio in 20 microliter of the amniotic fluids obtained at delivery, and the amniotic fluids from the women who delivered a baby suffering from respiratory distress syndrome (RDS) were easily discriminated from the normal amniotic fluids. In an analysis of 200 microliter of amniotic fluids from 4 RDS cases and 16 normal baby cases, the mean DSPC/Sm ratios were 0.97 +/- 0.53 and 5.75 +/- 1.29, respectively.
通过用单克隆抗体VJ - 41进行薄层层析免疫染色，该抗体优先与鞘磷脂（Sm）和含二饱和脂肪酸的磷脂酰胆碱（DSPC）发生反应，仅在人羊水的脂质提取物中检测到Sm和表面活性物质磷脂二棕榈酰磷脂酰胆碱（DPPC）。该方法对于选择性和同时测定羊水中表面活性物质磷脂和Sm的浓度以确定胎儿肺部成熟度水平很有用。通过测量薄层层析免疫染色显示的斑点密度，我们使用该抗体检测到Sm的灵敏度比检测DPPC的灵敏度高两倍。在同一薄层层析板上检测到超过50 ng的DPPC和Sm，标准曲线在磷脂含量达1微克时呈线性。该方法用于测定分娩时获得的20微升羊水中表面活性物质磷脂/Sm比值，患有呼吸窘迫综合征（RDS）婴儿的产妇的羊水很容易与正常羊水区分开来。在对4例RDS病例和16例正常婴儿病例的200微升羊水进行分析时，平均DSPC/Sm比值分别为0.97±0.53和5.75±1.29。
