Adaptability Research of HL60-pNL3.2 Reporter Gene Assay in Pyrogen Detection of Vaccines
10.13748/j.cnki.issn1007-7693.20222350
- VernacularTitle:HL60-pNL3.2报告基因热原检查法测定疫苗热原的适用性研究
- Author:
Can WANG
1
;
Lizhen LIU
1
;
Gang CHEN
1
;
Hong SHAO
1
Author Information
1. Shanghai Institute for Food and Drug Control, NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies, Shanghai 201203, China
- Publication Type:Journal Article
- Keywords:
pyrogen detection;
reporter gene assay;
live attenuated varicella vaccine;
inactivated influenza vaccine;
lyophilized live attenuated hepatitis A vaccine
- From:
Chinese Journal of Modern Applied Pharmacy
2024;41(3):354-358
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE :To explore the application of HL60-pNL3.2 reporter gene assay in pyrogen detection of live attenuated varicella vaccine, inactivated influenza vaccine and lyophilized live attenuated hepatitis A vaccine.
METHODS
According to the publication draft named as "in vitro pyrogen test(reporter gene assay)" in Chinese Pharmacopoeia, maximum valid dilution(MVD) of three kinds of vaccines was calculated, and the interference test was conducted to explore whether these vaccines interfere with the pyrogen determination. According to the general rule 9101 from Chinese Pharmacopoeia 2020, the assay in the pyrogen detection of the three vaccines was validated, using the methods to determine the pyrogen content of three vaccines.
RESULTS
The MVD of live attenuated varicella vaccine, inactivated influenza vaccine and lyophilized live attenuated hepatitis A vaccine respectively were 1 000, 200 and 1 000 times. The recovery rates of endotoxin(LPS) in the three kinds of vaccine were between 52.5% and 110.1%. The methodological validation results showed that the method could be used for the determination of pyrogens in three vaccines, and the correlation coefficient(R2) between LPS concentration and its chemical light intensity(RLU value) was > 0.98, the recovery rates were between 76.4% and 192.8% when LPS concentration was between 1 and 250 EU·mL–1. The coefficient of variation of measured results was less than 25% when LPS concentration was more than 5 EU·mL–1. The limit of detection and the limit of quantitation were between 0.05 and 0.13 EU·mL–1. Then, the pyrogen of these vaccines was determined by HL60-pNL3.2 reporter gene assay, and the pyrogen concentration all was less than contaminant limit concentration.
CONCLUSION
HL60-pNL3.2 reporter gene assay has the advantages of no animal, simple and rapid operation, with wide pyrogen spectrum and can quantitative pyrogen, which can be used to detect pyrogen of live attenuated varicella vaccine, inactivated influenza vaccine and lyophilized live attenuated hepatitis A vaccine.
