Considerations and Some Practical Solutions to Overcome Nanoparticle Interference with LAL Assays and to Avoid Endotoxin Contamination in Nanoformulations
Monitoring endotoxin contamination in drugs and medical devices is required to avoid pyrogenic response and septic shock in patients receiving these products. Endotoxin contamination of engineered nanomaterials and nanotechnology-based medical products represents a significant translational hurdle. Nanoparticles often interfere with an in vitro Limulus Amebocyte Lysate (LAL) assay commonly used in the pharmaceutical industry for the detection and quantification of endotoxin. Such interference challenges the preclinical development of nanotechnology-formulated drugs and medical devices containing engineered nanomaterials. Protocols for analysis of nanoparticles using LAL assays have been reported before. Here, we discuss considerations for selecting an LAL format and describe a few experimental approaches for overcoming nanoparticle interference with the LAL assays to obtain more accurate estimation of endotoxin contamination in nanotechnology-based products. The discussed approaches do not solve all types of nanoparticle interference with the LAL assays but could be used as a starting point to address the problem. This chapter also describes approaches to prevent endotoxin contamination in nanotechnology-formulated products.
Key wordsEndotoxin LAL Interference Inhibition enhancement control
This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.
- 1.Brade H, Opal SM, Vogel SN, Morrison DC (eds) (1999) Endotoxin in health and disease. Marcel Dekker Inc., New YorkGoogle Scholar
- 3.USP 30 NF 25 (2007) <85> Bacterial endotoxins test. vol 1Google Scholar
- 4.HHS, US FDA (2012) Guidance for industry. Pyrogen and endotoxins testing: questions and answers. https://www.fda.gov/downloads/drugs/guidances/ucm310098.pdf
- 5.HHS, US FDA (2015) .Guidance for Industry and Food and Drug Administration Staff. Endotoxin testing recommendations for single-use intraocular ophthalmic devices. https://www.fda.gov/downloads/medicaldevices/deviceregulationandguidance/guidancedocuments/ucm393376.pdf
- 21.Lyons JL, Roos KL, Marr KA, Neumann H, Trivedi JB, Kimbrough DJ, Steiner L, Thakur KT, Harrison DM, Zhang SX (2013) Cerebrospinal fluid (1,3)-beta-D-glucan detection as an aid for diagnosis of iatrogenic fungal meningitis. J Clin Microbiol 51(4):1285–1287. doi: 10.1128/jcm.00061-13 CrossRefPubMedPubMedCentralGoogle Scholar
- 25.Sandle T (2011) A practical approach to depyrogenation studies using bacterial endotoxin. J GxP Compliance 15(4):90–96Google Scholar
- 27.Zheng J, Clogston JD, Patri AK, Dobrovolskaia MA, McNeil SE (2011) Sterilization of silver nanoparticles using standard gamma irradiation procedure affects particle integrity and biocompatibility. J Nanomed Nanotechnol 2011(Suppl 5):001. doi: 10.4172/2157-7439.s5-001 PubMedPubMedCentralGoogle Scholar