Further purification of epidermal growth factor by high-performance liquid chromatography.
Further purification of epidermal growth factor by high-performance liquid chromatography.
复制标题
采用高效液相色谱法进一步纯化表皮生长因子。
DOI：
10.1016/0003-2697(82)90015-x
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发表时间：
1982
影响因子：
2.9
通讯作者：
Magun,BE
中科院分区：
文献类型：
--
作者：
Matrisian,LM;Larsen,BR;Finch,JS;Magun,BE
通过Savage和Cohen的方法（J. Biol. Chem. 247，7601-7611（1972），使用DEAE-纤维素色谱作为最终纯化步骤纯化的表皮生长因子（EGF）通过反相高效液相色谱（HPLC）进一步分解成两种主要的紫外吸收组分。这两种成分，称为α-EGF和β-EGF，与125 I-标记的EGF竞争EGF受体，诱导新生小鼠过早睁眼，并且具有与Savage等（J. Biol. Chem. 247，7612-7621（1972））公开的氨基酸组成相似的氨基酸组成。在十二烷基硫酸钠-尿素-聚丙烯酰胺凝胶电泳中，β-EGF的迁移速度略快于α-EGF。α-EGF刺激静止期Rat-1细胞DNA合成的能力比β-EGF强4倍，比DEAE纯化的EGF强10倍。EGF的HPLC纯化可以取代目前使用的DEAE-纤维素色谱步骤，并产生更有效和更少的异质性EGF物种。
Epidermal growth factor (EGF) purified by the method of Savage and Cohen (J. Biol. Chem.247, 7601–7611 (1972) using DEAE-cellulose chromatography as the final purification step was further resolved into two major uv-absorbing components by reverse-phase high-performance liquid chromatography (HPLC). Both components, referred to as α-EGF and β-EGF, competed with125I-labeled EGF for the EGF receptor, induced premature eye opening in neonatal mice, and had an amino acid composition similar to that published by Savage et al. (J. Biol. Chem.247, 7612–7621 (1972). β-EGF migrated slightly faster than α-EGF during sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis. α-EGF was fourfold more potent than β-EGF and was 10-fold more potent than DEAE-purified EGF in stimulating DNA synthesis in quiescent Rat-1 cells. HPLC purification of EGF can replace the DEAE-cellulose chromatography step currently used and produces a more potent and less heterogeneous EGF species.