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Bioscience Methods
BM 2011, Vo.2, No.1
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Figure 7 HPLC chromatogram of NGF standard
Note: The chromatogram reveals the exclusive NGF peak and suggests that the purity of standard is 99.1% from statistics
Figure 8 Mass spectrogram of NGF standard
Note: It reveals that mainly exists the NGF; The three peak
groups represent the NGF double-charged peak, single-charged
peak and dimer peak. Meanwhile, the doublet of every peak
group reveals the 8 amino acids dissimilarity from the
post-transcriptional modification
3.3 Method
In order to prevent the protein denaturation and
bringing in impurities, we chose SEC-HPLC other
than the traditional RP-HPLC method which will
introduce in impurities such as acetonitrile and
methanol. This work used Shimadzu Shodex
KW-802.5 column that exclusion range of 6~15 kD.
Author Contributions
Da Huang and Hongwei Ren designed research; Da Huang and
Jianrong Zhang performed research; Da Huang analyzed data;
Da Huang wrote the paper; and Yiming Xu, Liming Lu,
Lingyan Ma, Yongchao Fu, Minxiang Li and Lingyuan
Xiong provided support data and advisory. The authors declare
no conflict of interest.
Acknowledgements
This work was supported by the Xiamen Bioway Biotech Co.
Ltd., and the School of Life Science, Peking University, China.
References
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William C.M., Schenker A., Shooter E.M., 1976, Characterization and
isolation of proteolytically modified nerve growth factor, Biochemistry,
15(25): 5543-5552