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A reporter gene assays for bioactivity determination of human chorinonic gonadotropin
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Ying HUANG1, 2, Xiao-ming ZHANG2, He-yang LI2, Lü-yin WANG2, Hui ZHANG2, Ping LÜ2, Jing LI2, *, Xiang-dong GAO1, *, Cheng-gang LIANG2, *
Acta Pharmaceutica Sinica | 2024, 59(2) : 432 - 438
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Acta Pharmaceutica Sinica | 2024, 59(2): 432-438
• Original Articles •
A reporter gene assays for bioactivity determination of human chorinonic gonadotropin
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Ying HUANG1, 2, Xiao-ming ZHANG2, He-yang LI2, Lü-yin WANG2, Hui ZHANG2, Ping LÜ2, Jing LI2, *, Xiang-dong GAO1, *, Cheng-gang LIANG2, *
Affiliations
  • 1. China Pharmaceutical University, Nanjing 211198, China
  • 2. National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Beijing 102629, China
Published: 2024-02-12 doi: 10.16438/j.0513-4870.2023-0545
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This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control.

reporter gene assay  /  human chorinonic gonadotropin  /  four-parameter curve  /  biological activity  /  method validation
Ying HUANG, Xiao-ming ZHANG, He-yang LI, Lü-yin WANG, Hui ZHANG, Ping LÜ, Jing LI, Xiang-dong GAO, Cheng-gang LIANG. A reporter gene assays for bioactivity determination of human chorinonic gonadotropin[J]. Acta Pharmaceutica Sinica, 2024 , 59 (2) : 432 -438 . DOI: 10.16438/j.0513-4870.2023-0545
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Year 2024 volume 59 Issue 2
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Article Info
doi: 10.16438/j.0513-4870.2023-0545
  • Receive Date：2023-04-28
  • Online Date：2025-11-28
  • Published：2024-02-12
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History
  • Received：2023-04-28
  • Revised：2023-09-08
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Affiliations
    1. China Pharmaceutical University, Nanjing 211198, China
    2. National Institutes for Food and Drug Control, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, Beijing 102629, China
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表12种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
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Genus		 种数
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Percentage of total
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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