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中国生物工程杂志

China Biotechnology
China Biotechnology  2016, Vol. 36 Issue (6): 76-80    DOI: 10.13523/j.cb.20160610
    
Measurement and Analysis of Recombinant Basic Fibroblast Growth Factor's Free Sulfhydryl
DENG Chun-pin, YANG Bo, MEI Xiong, ZHENG Zan-shun, QU Wei
GuangDong Engineering Research Center of Cell Factor, Zhuhai ESSEX Bio-Pharmaceutical Company Limited, Zhuhai 519020, China
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Abstract  

A rapid and simple analysis method based on uv-vis spectrophotometry technology was established to measure the concentration of free sulfhydryl groups in recombinant basic fibroblast growth factor. Detection reagents,standard references and samples were mixed, incubated for some time,followed by OD412 value reading with uv-vis spectrometer. Standard curve was generated by taking the concentration of standard reference as the horizontal coordinate, taking the OD412 values as the vertical coordinate. The concentration of free sulfhydryl groups in tested samples was calculated according to standard curve. The method qualification results showed that the method is free sulfhydryl specific, with accuracy range 91.7%~107.4%, precision range 5%~8%, and linearity range 20~80μg/ml. The measured result of free sulfhydryl groups on molecular surface or total were consistent with the theoretical value.This method can be used to measure the free sulfhydryl groups in proteins to monitor the correctness and homogeneity of molecular structure during the quality research of related biopharmaceuticals development.



Key wordsSulfhydryl      Ellman's method      Basic fibroblast growth factor     
Received: 09 December 2015      Published: 26 January 2016
ZTFLH:  Q819  
Cite this article:

DENG Chun-pin, YANG Bo, MEI Xiong, ZHENG Zan-shun, QU Wei. Measurement and Analysis of Recombinant Basic Fibroblast Growth Factor's Free Sulfhydryl. China Biotechnology, 2016, 36(6): 76-80.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20160610     OR     https://manu60.magtech.com.cn/biotech/Y2016/V36/I6/76

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