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Open AccessArticle

Bioactivity Determination of a Therapeutic Recombinant Human Keratinocyte Growth Factor by a Validated Cell-based Bioassay

by Wenrong Yao 1,2,†, Ying Guo 1,†, Xi Qin 1,†, Lei Yu 1, Xinchang Shi 1, Lan Liu 1, Yong Zhou 1, Jinpan Hu 1, Chunming Rao 1,* and Junzhi Wang 1,*
National Institutes for Food and Drug Control, No. 2, Tiantan Xili, Dongcheng District, Beijing 100050, China
Institutes of Process Engineering, Chinese Academy of Sciences, 1 North 2nd Street, Zhongguancun, Haidian District, Beijing 100190, China
Authors to whom correspondence should be addressed.
They contributed equally to this work.
Academic Editors: In-Soo Yoon and Hyun-Jong Cho
Molecules 2019, 24(4), 699;
Received: 19 January 2019 / Revised: 12 February 2019 / Accepted: 13 February 2019 / Published: 15 February 2019
(This article belongs to the Special Issue Method Development and Validation in Food and Pharmaceutical Analysis)
The therapeutic recombinant human keratinocyte growth factor 1 (rhKGF-1) was approved by the FDA for oral mucositis resulting from hematopoietic stem cell transplantation for hematological malignancies in 2004. However, no recommended bioassay for rhKGF-1 bioactivity has been recorded in the U.S. Pharmacopoeia. In this study, we developed an rhKGF-1-dependent bioassay for determining rhKGF-1 bioactivity based on HEK293 and HaCat cell lines that stably expressed the luciferase reporter driven by the serum response element (SRE) and human fibroblast growth factor receptor (FGFR2) IIIb. A good responsiveness to rhKGF-1 and rhKGF-2 shared by target HEK293/HaCat cell lines was demonstrated. Our stringent validation was completely focused on specificity, linearity, accuracy, precision, and robustness according to the International Council for Harmonization (ICH) Q2 (R1) guidelines, AAPS/FDA Bioanalytical Workshop and the Chinese Pharmacopoeia. We confirmed the reliability of the method in determining rhKGF bioactivity. The validated method is highly timesaving, sensitive, and simple, and is especially valuable for providing information for quality control during the manufacture, research, and development of therapeutic rhKGF. View Full-Text
Keywords: rhKGF-1; rhKGF-2; bioactivity; cell-based bioassay; method validation rhKGF-1; rhKGF-2; bioactivity; cell-based bioassay; method validation
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MDPI and ACS Style

Yao, W.; Guo, Y.; Qin, X.; Yu, L.; Shi, X.; Liu, L.; Zhou, Y.; Hu, J.; Rao, C.; Wang, J. Bioactivity Determination of a Therapeutic Recombinant Human Keratinocyte Growth Factor by a Validated Cell-based Bioassay. Molecules 2019, 24, 699.

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