Separation of neuropeptide Y diastereomers by high-performance liquid chromatography and capillary zone electrophoresis.
Separation of neuropeptide Y diastereomers by high-performance liquid chromatography and capillary zone electrophoresis.
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通过高效液相色谱和毛细管区带电泳分离神经肽 Y 非对映体。
DOI：
10.1016/0021-9673(93)83309-g
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Rivier,JE
中科院分区：
文献类型：
--
作者：
Kirby,DA;Miller,CL;Rivier,JE
分离类似物的神经肽Y（NPY），其中一个单一的氨基酸取代相应的天然存在的残基进行色谱技术，以确保质量的合成肽用于结构和生物学研究。在35种化合物中，28种通过标准反相高效色谱（RP-HPLC）方法容易地从天然NPY中分离（α = 1.02-2.76），所述方法使用在我们实验室中开发的Vapriac C18柱和梯度缓冲系统，所述梯度缓冲系统由磷酸三乙铵（TEAP）（pH 2.25）和乙腈（40°C）组成。相同的非对映异构体可以在相同的固体载体上分离，并使用0.1%的三氟乙酸（TFA）作为移动的相改性剂，但分离因子较小，保留时间较长。通过将固相支持物更换为Vaporac二苯基衍生化硅胶和由0.1% TFA和乙腈组成的缓冲液系统，分离了剩余7种未分离类似物中的3种（α = 1.02-1.96）。在剩下的四个未解析的类似物中，只有两个可以通过毛细管区带电泳（CZE）在pH 2.5的0.1M磷酸钠中分离，但所有四个最终通过将电泳缓冲液改为pH 2.5的0.1MTEAP缓冲液来解析。非对映体的迁移时间与天然NPY的迁移时间相差0.2-2.0 min。除了确认每种异构体的独特性外，本研究还证明了TEAP缓冲系统对RP-HPLC和CZE的广泛实用性和高效性，以及TEAP和TFA缓冲液在RP-HPLC中产生的选择性差异。这里描述的条件具有广泛的应用，用于合成和天然肽的分析和制备分离。
Separation of analogues of neuropeptide Y (NPY) in which a singled-amino acid replaced the corresponding naturally occurring residue was performed by chromatographic techniques to ensure the quality of the synthetic peptides to be used for structural and biological studies. Of the 35 compounds, 28 were easily separated (α = 1.02–2.76) from native NPY by standard reversed-phase high-performance chromatography (RP-HPLC) methods using a Vydac C18column and a gradient buffer system developed in our laboratory comprised of triethylammonium phosphate (TEAP) at pH 2.25 and acetonitrile at 40°C. The identical diastereomers could be separated on the same solid support and by using 0.1% trifluoroacetic acid (TFA) as the mobile phase modifier, however separation factors were smaller and retention times were longer. Three of the remaining seven unresolved analogues were separated (α = 1.02–1.96) by changing the solid-phase support to Vydac diphenyl derivatized silica and a buffer system consisting of 0.1% TFA and acetonitrile. Of the four remaining unresolved analogues, only two could be separated by capillary zone electrophoresis (CZE) in 0.1Msodium phosphate at pH 2.5, but all four were finally resolved by changing the electrophoretic buffer to 0.1MTEAP buffer at pH 2.5. Migration times of the diastereomers differed by 0.2–2.0 min from that of the natural NPY. In addition to confirming the uniqueness of each isomer, this investigation demonstrated the expansive utility and high efficiency of the TEAP buffer system for both RP-HPLC and CZE as well as the difference in selectivity produced by the TEAP and TFA buffers in RP-HPLC. The conditions described here have broad applications for the analysis and preparative separation of synthetic and native peptides.
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D-Trp8-生长抑素：生长抑素的类似物，比天然分子更有效。
DOI：
--
发表时间：
1975
期刊：
Biochemical and Biophysical Research Communications - BBRC
影响因子：
--
作者：
J. Rivier;M. Brown;W. Vale
通讯作者：
W. Vale
影响因子：
2.9
作者：
P. D. Grossman;K. Wilson;G. Petrie;H. Lauer
通讯作者：
P. D. Grossman;K. Wilson;G. Petrie;H. Lauer
DOI：
10.1111/j.1399-3011.1989.tb00677.x
发表时间：
1989
期刊：
International journal of peptide and protein research
影响因子：
--
作者：
Boublik,J;Scott,N;Taulane,J;Goodman,M;Brown,M;Rivier,J
通讯作者：
Rivier,J
β-内啡肽的反相色谱行为：构象变化的证据。
影响因子：
4.1
作者：
L. Monger;C. Olliff
通讯作者：
C. Olliff
在反相 HPLC 中使用三烷基磷酸铵 (TAAP) 缓冲液实现肽和蛋白质的高分辨率和高回收率
DOI：
--
发表时间：
1978
期刊：
影响因子：
--
作者：
J. Rivier
通讯作者：
J. Rivier