Research Progress in Detection Methods of Pyrogen and Bacterial Endotoxin

doi:10.19803/j.1672-8629.20240821

Abstract

Abstract: Objective To introduce the detection methods of pyrogen and bacterial endotoxin in order to provide a reference for establishing a standardized detection system. Methods The characteristics, research progress and developments related to pyrogen detection in vivo, pyrogen detection in vitro and other detection methods were outlined based on related literature. Results The rabbit pyrogen method was being phased out globally while non-animal pyrogen and bacterial endotoxin detection methods became more popular. However, each in vivo method had its own technical limitations. Conclusion The limitations of each in vivo detection method make it necessary to ensure the safety of drugs for injection and medical devices.

Key words: Pyrogen, Bacterial Endotoxin, Limulus Reagent, Rabbit Pyrogen Test, Cellular Method, Recombinant Factor C, Detection Method, Alternative Method

CLC Number:

R927
R994.11

LIU Sihan, PEI Yusheng, TAN Dejiang, SUN Huimin, ZHANG Heng, LIU Tao, WANG Dongmei, HE Qing. Research Progress in Detection Methods of Pyrogen and Bacterial Endotoxin[J]. Chinese Journal of Pharmacovigilance, 2025, 22(4): 361-367.

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URL: https://zgywjj.magtechjournal.com/EN/10.19803/j.1672-8629.20240821

https://zgywjj.magtechjournal.com/EN/Y2025/V22/I4/361

References

[1] KELLUM JA, RONCO C.The Role of Endotoxin in Septic Shock[J]. Critical Care, 2023, 27(1): 400.
[2] ŚMIECHOWICZ J.The Rationale and Current Status of Endotoxin Adsorption in the Treatment of Septic Shock[J]. Journal of Clinical Medicine, 2022, 11(3): 619.
[3] KOCIJANCIC D, LESCHNER S, FELGNER S, et al.Therapeutic Benefit of Salmonella Attributed to LPS and TNF-α is Exhaustible and Dictated by Tumor Susceptibility[J]. Oncotarget, 2017, 8(22): 36492.
[4] ZHANG LL, CHEN B, FAN XY, et al.LPS Cooperates with Poly-L-Arginine to Promote IL-6 and IL-8 Release via the JNK Signaling Pathway in NCI-H292 Cells[J]. Journal of Immunology Research, 2016, 2016(1): 3421060.
[5] SONDHI P, ADENIJI T, LINGDEN D, et al.Advances in Endotoxin Analysis[J]. Advances in Clinical Chemistry, 2024, 118: 1-34.
[6] KIM J H, JUNG K, KIM J, et al.Development of a Rabbit Monocyte Activation Test as An Alternative to the Rabbit Pyrogen Test and Its Application in the Analysis of Plasma-Derived Products[J]. Biologicals, 2021, 71: 20-30.
[7] CIREFICE G, SCHÜTTE K, SPREITZER I, et al. The Future of Pyrogenicity Testing: Phasing out the Rabbit Pyrogen Test, A Meeting Report[J]. Biologicals, 2023, 84: 101702.
[8] European Pharmacopoeia Commission. European Pharmacopoeia Bids Adieu to Rabbit Pyrogen Test in Its Monographs[EB/OL]. (2024-06-16)[2024-10-11]. https://www.edqm.eu/en/-/european-pharm-acopoeia-to-put-an-end-to-the-rabbit-pyrogen-test.
[9] ABDIN SM, MANSEL F, HASHTCHIN AR, et al.Sensor Macrophages Derived from Human Induced Pluripotent Stem Cells to Assess Pyrogenic Contaminations in Parenteral Drugs[J]. Biofabrication, 2024, 16(3): 035017.
[10] Chinese Pharmacopoeia Commission.Pharmacopoeia of the People's Republic of China, Volume IV(中华人民共和国药典四部)[M]. Beijing: China Medical Science Press, 2020: 78-181, 474, 515-517.
[11] WANG C, DONG S, ZHAO H, et al.A Novel Monocyte-based Pyrogen Test Based on the Mechanism of Human Fever Reaction[J]. Current Pharmaceutical Analysis, 2016, 12(3): 227-233.
[12] ZHANG Y, CAI T, DU Y, et al.Suitability of Human Whole Blood in vitro Pyrogen Test to Two Vaccines[J]. Chinese Journal of Biologicals(中国生物制品学杂志), 2018, 31(9): 995-998.
[13] WANG C,XU L,HUA XD.The Study on HL-60/IL-6 in vitro Pyrogen Detection Method and Its Applicability to Cell Therapy Products[J]. Tianjin Pharmacy(天津药学), 2020, 32(2): 6-10.
[14] PEI YS, CHEN C, NING X, et al.Establishment and Application of the First Batch of Nation Astandards for Bacterial Endotoxin for Monocyte Activation Reaction Tests[J]. Chinese Journal of Pharmac-15 ovigilance(中国药物警戒), 2023, 20(11): 1217-1220.
[15] MOLENAAR-DE BACKER MWA, DOODEMAN P, REZAI F, et al. In vitro Alternative for Reactogenicity Assessment of Outer Membrane Vesicle Based Vaccines[J]. Scientific Reports, 2023, 13(1): 12675.
[16] WANG C, WANG ZQ, CAO Y, et al.Interpretation of the Draft "in vitro Pyrogen Test (Reportergene Assay)" for the Chinese Pharmacopoeia[J]. Drug Standards of China(中国药品标准), 2022, 23(2): 148-151.
[17] LI K, XU H, JIANG W, et al.Development and Characterization of Stable Reporter Cells for Fast and Sensitive Detection of Pyrogen[J]. Analytical Biochemistry, 2018, 557: 69-76.
[18] HE Q, YU C, WANG L, et al.A Novel Reporter Gene Assay for Pyrogen Detection[J]. Japanese Journal of Infectious Diseases, 2020, 73(2): 111-118.
[19] WANG C, WANG M, LIU L, et al.Development and Validation of a Novel Luciferase Reporter Gene Assay to Detect Pyrogen[J]. Biologicals, 2022, 77: 16-23.
[20] HE Q, YU CF, WU G, et al.A novel Alternative for Pyrogen Detection Based on a Transgenic Cell Line[J]. Signal Transduction and Targeted Therapy, 2024, 9(1): 33.
[21] HAN Q, HU R, LI H, et al.Application of a TLR over Expression Cell Model in Pyrogen Detection[J]. Biotechnology and Bioengineering, 2019, 116(6): 1269-1279.
[22] HU R, LI H, LEI Z, et al.Construction of a Sensitive Pyrogen-Testing Cell Model by Site-Specific Knock-In of Multiple Genes[J]. Biotechnology and Bioengineering, 2019, 116(10): 2652-2661.
[23] CHEN M, WANG C, WANG W, et al.De Novo Assembly and Characterization of Early Embryonic Transcriptome of the Horseshoe Crab Tachypleus Tridentatus[J]. PLoS One, 2016, 11(1): e0145825.
[24] BANG FB.A Bacterial Disease of Limulus Polyphemus[J]. Bull Johns Hopkins Hosp, 1956, 98(5): 325-351.
[25] MUTA T, IWANAGA S.Clotting and Immune Defense in Limulidae[J]. Invertebrate Immunology, 1996, 15: 154-189.
[26] ABATE W, SATTAR AA, LIU J, et al.Evaluation of Recombinant Factor C Assay for the Detection of Divergent Lipopolysaccharide Structural Species and Comparison with Limulus Amebocyte Lysate-Based Assays and a Human Monocyte Activity Assay[J]. Journal of Medical Microbiology, 2017, 66(7): 888-897.
[27] DING JL, HO B.Endotoxin Detection-from Limulus Amebocyte Lysate to Recombinant Factor C[J]. Endotoxins: Structure, Function and Recognition, 2010, 53: 187-208.
[28] KOBAYASHI M, MORIMOTO T, YAMAMOTO M.Studies on the Gelation Reaction of Limulus Lysate (Pre-Gel). Iii. Influences Of Various Substances on the Gelation Reaction of Limulus Lysate (Author's Transl)[J]. Yakugaku Zasshi: Journal of the Pharmaceutical Society of Japan, 1975, 95(6): 720-727.
[29] KWAN KY, BOPP J, HUANG S, et al.Ontogenetic Resource Use and Trophic Dynamics of Endangered Juvenile Tachypleus Tridentatus Among Diversified Nursery Habitats in the Northern Beibu Gulf, China[J]. Integrative Zoology, 2021, 16(6): 908-928.
[30] BAKER E, PONDER J, OBERDORFER J, et al.Barriers to the Use of Recombinant Bacterial Endotoxins Test Methods in Parenteral Drug, Vaccine and Device Safety Testing[J]. Alternatives to Laboratory Animals, 2023, 51(6): 401-410.
[31] GUO B. Protecting the "Living Marine Fossil" Chinese Horseshoe Crab[N]. Fujian Daily(福建日报), 2023-11-17(008).
[32] ANDERSON RL, WATSON III WH, CHABOT CC.Sublethal Behavioral and Physiological Effects of the Biomedical Bleeding Process on the American Horseshoe Crab, Limulus Polyphemus[J]. The Biological Bulletin, 2013, 225(3): 137-151.
[33] MCKENZIE JH, ALWIS KU, SORDILLO JE, et al.Evaluation of Lot-to-Lot Repeatability and Effect of Assay Media Choice in the Recombinant Factor C Assay[J]. Journal of Environmental Monitoring, 2011, 13(6): 1739-1745.
[34] TINDALL B, DEMIRCIOGLU D, UHLIG T.Recombinant Bacterial Endotoxin Testing: a Proven Solution[J]. BioTechniques, 2021, 70(5): 290-300.
[35] SU W, DING X.Methods of Endotoxin Detection[J]. Journal of Laboratory Automation, 2015, 20(4): 354-364.
[36] DING JL, HO B.A New Era in Pyrogen Testing[J]. Trends in Biote-chnology, 2001, 19(8): 277-281.
[37] MIZUMURA H, OGURA N, AKETAGAWA J, et al.Genetic Engineering Approach to Develop Next-Generation Reagents for Endotoxin Quantification[J]. Innate Immunity, 2017, 23(2): 136-146.
[38] ABATE W, SATTAR AA, LIU J, et al.Evaluation of Recombinant Factor C Assay for the Detection of Divergent Lipopolysaccharide Structural Species and Comparison with Limulus Amebocyte Lysate-Based Assays and a Human Monocyte Activity Assay[J]. Journal of Medical Microbiology, 2017, 66(7): 888-897.
[39] BOLDEN J, SMITH K.Application of Recombinant Factor C Reagent for the Detection of Bacterial Endotoxins in Pharmaceutical Products[J]. PDA Journal of Pharmaceutical Science and Technology, 2017, 71(5): 405-412.
[40] YANG SY, WEN ZT, RONG DY, et al.Research Progress of Recombinant Limulus C Factor (rFC) Replacing Limulus Blood in Preparation of Limulus Reagents[J]. Heilongjiang Science(黑龙江科学), 2024, 15(8): 118-120.
[41] SUI X, YE XY, HUANG QY, et al.Suggestion on Application of Recombinant C Factor Bacteria Endotoxion Assay to Medica Devices[J]. China Medical Device Information(中国医疗器械信息), 2023, 29(7): 27-29, 39.
[42] USP Microbiology Expert Committee. Expert Committee Approves Endotoxin Testing Using Non-Animal Derived Reagents[EB/OL]. (2024-07-26) [2024-10-11]. https://www.usp.org/news/expert-committee-approves-endotoxin-testing-using-non-animal-derived-reagents.
[43] MARIUS M, VACHER F, BONNEVAY T.Comparison of LAL and Recombinant Factor C Endotoxin Testing Assays in Human Vaccines with Complex Matrices[J]. PDA Journal of Pharmaceutical Science and Technology, 2020, 74(4): 394-407.
[44] KANG DH, YUN SY, EUM SY, et al.A Study on the Application of Recombinant Factor C (rFC) Assay Using Biopharmaceuticals[J]. Microorganisms, 2024, 12(3): 516.
[45] MUROI M, OGURA N, MIZUMURA H, et al.Application of a Recombinant Three-Factor Chromogenic Reagent, Pyrosmart, for Bacterial Endotoxins Test Filed in the Pharmacopeias[J]. Biological and Pharmaceutical Bulletin, 2019, 42(12): 2024-2037.
[46] LIU SX, CHEN T, RUAN J.Suitability Study of Recombinant Limulus Amebocyte Lysate for the Determination of Bacterial Endotoxins in Hepatocyte Growth-Promoting Factors Injection[J]. China Pharmaceuticals(中国药业), 2023, 32(22): 92-95.
[47] CHEN X, HE LM, LIU R.Progress on Detection of Bacterial Endot-oxin[J]. Chinese Journal of Analysis Laboratory(分析试验室), 202348 42(9): 1242-1250.
[48] FUNG FM, SU M, FENG H, et al.Extraction, Separation and Characterization of Endotoxins in Water Samples Using Solid Phase Extraction and Capillary Electrophoresis-Laser Induced Fluorescence[J]. Scientific Reports, 2017, 7(1): 10774.
[49] GIANNAKOU C, AIMONEN K, BLOOIS L, et al.Sensitive Method for Endotoxin Determination in Nanomedicinal Product Samples[J]. Nanomedicine, 2019, 14(10): 1231-1246.
[50] HONG G, HAN YK, YEO MK, et al.Development of Analytical Methods for the Determination of 3-Hydroxy Fatty Acids and Muramic Acid as Bacterial Markers in Airborne Particles and Settled Dust[J]. Journal of Chromatography A, 2023, 1688: 463708.
[51] ZHANG Y, SUN SX.Research Progress of Bacterial Endotoxin Detection Methods[J]. Chinese Journal of Biologicals(中国生物制品学杂志), 2023, 36(3): 368-372, 378.
[52] WANG S, SHEN F, HE XM, et al.Application of Biosensors in the Detection of Escherichia Coli Endotoxin[J]. Journal of Food Safety & Quality(食品安全质量检测学报), 2023, 14(13): 48-55.
[53] AKIMOV N, SCUDDER J, YE JY.Refinement of an Open-Microcavity Optical Biosensor for Bacterial Endotoxin test[J]. Biosensors and Bioelectronics, 2021, 191: 113436.
[54] CUI Q, LI J, LI Y, et al.Sensitive and Rapid Detection of Bacterial Endotoxin with a Functional Carbon Nanotube Field-Effect Transistor Biosensor[J]. Talanta, 2024, 266: 125035.
[55] WANG S, YAN Z, SHEN F, et al.Novel Aptasensor Based on Polyaniline Functionalized Carboxylated Dobby Carbon Nanotubes and Molybdenum Disulfide for Endotoxin Detection[J]. Talanta, 2024, 276: 126256.
[56] HE Q, TAN DJ, MA X, et al.Progress and Challenges in the Implementation of 3Rs Qualitycontrol Approaches in WHO Guidelines on Biologicals[J]. Chinese Journal of New Drugs(中国新药杂志), 2024, 33(16): 1659-1664.
[57] ZHANG C, LIU WJ, ZHANG ML, et al.Review on Characteristics and Detecting Assay of Bacterial Endotoxin Contamination in Water Envir-onment[J]. Environmental Science(环境科学), 2014, 35(4): 1597-1601.