Youn Yu Seok, Na Dong Hee, Yoo Sun Dong, Song Soo-Chang, Lee Kang Choon
Drug Targeting Laboratory, College of Pharmacy, SungKyunKwan University, 300 Chonchon-dong, Jangan-ku, Suwon City 440-746, South Korea.
J Chromatogr A. 2004 Dec 17;1061(1):45-9. doi: 10.1016/j.chroma.2004.10.062.
A one-step chromatographic method capable of separating all isomers of polyethylene glycol (PEG)-growth hormone-releasing factor (GRF) (1-29) conjugates was developed. The unmodified GRF (1-29) and seven different isomers of PEG-GRF (1-29) conjugates were separated by using a simple reversed-phase HPLC method depending on the differences of hydrophobicity due to the number and site of PEG attachment. The PEGylation sites of all isomers of PEG-GRF (1-29) conjugates were identified by determining the molecular masses of the Lys-C digested fragments with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This study is a first report for the separation of all PEG-conjugate isomers and would be useful for further studies to find the promising conjugate by evaluating biological activity and stability of each isomer.
开发了一种能够分离聚乙二醇（PEG）-生长激素释放因子（GRF）（1-29）缀合物所有异构体的一步色谱法。未修饰的GRF（1-29）和PEG-GRF（1-29）缀合物的七种不同异构体通过一种简单的反相高效液相色谱法进行分离，该方法基于PEG连接数量和位点导致的疏水性差异。通过基质辅助激光解吸/电离飞行时间质谱法测定Lys-C消化片段的分子量，确定了PEG-GRF（1-29）缀合物所有异构体的PEG化位点。本研究是关于分离所有PEG缀合物异构体的首次报道，对于通过评估每种异构体的生物活性和稳定性来寻找有前景的缀合物的进一步研究将是有用的。
