Quantitative Determination for Nerve Growth Factor Concentration in Commercial Product by SECâ€HPLC  

Da Huang1,2 , Jianrong Zhang1 , Yongchao Fu1 , Liming Lu2 , Yiming Xu1,2 , Lingyan Ma1 , Minxiang Li1 , Lingyuan Xiong1 , Hongwei Ren1,2
1. Xiamen Bioway Biotech Co. Ltd., Xiamen, 361009, P.R.China
2. Biochemistry & Molecular Biology College of Life Sciences, Peking University, Beijing, 100871, P.R. China
Xiamen Bioway Biotech Co. Ltd.
Author    Correspondence author
Bioscience Methods, 2011, Vol. 2, No. 1   doi: 10.5376/bm.2011.02.0001
Received: 23 Nov., 2010    Accepted: 12 Apr., 2011    Published: 18 Apr., 2011
© 2011 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Huang et al., 2011, Quantitative Determination for Nerve Growth Factor Concentration in Commercial Product by SEC-HPLC, Bioscience Methods, doi:10.5376/bm.2011.02.0001

Abstract

The commercial product, Mouse Nerve Growth Factor (NGF) for Injection-Nobex®, is consisted of NGF and human albumin. These two components are both proteins and make it difficult to determine the concentration of NGF by conventional methods. In order to establish a new efficiency quality control process for NGF, this work choose Size Exclusion High Performance Liquid Chromatography (SEC-HPLC) to detect stock solutions and products under screened chromatographic conditions: using Shodex KW-802.5 column, at room temperature with the 0.05 M PBS pH=7 buffer as mobile phase, 0.8 mL/min flow rate and detected under 280 nm wavelength. The standard curves with correlation coefficient of 0.999 82 (for stock solutions) and 0.999 97 (for products) were obtained by external standard method. The NGF concentrations tested in HPLC were compared with which in BCA for the same batches, and the RSD values were less than 3%, which corresponded with China Pharmacopoeia (2010). The results show that SEC-HPLC method is sensitive, accurate and reliable for NGF on-line quality control and has the potential for the quantitative determination of other mixed protein preparations.

Keywords
Size Exclusion High Performance Liquid Chromatography (SEC-HPLC); Mouse Nerve Growth Factor (NGF); Quantitative determination
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