Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Brade H, Opal SM, Vogel SN, Morrison DC (eds) (1999) Endotoxin in health and disease. Marcel Dekker Inc., New York
Dobrovolskaia MA, Vogel SN (2002) Toll receptors, CD14, and macrophage activation and deactivation by LPS. Microbes Infect 4(9):903–914. doi:10.1016/S1286-4579(02)01613-1
USP 30 NF 25 (2007) <85> Bacterial endotoxins test. vol 1
HHS, US FDA (2012) Guidance for industry. Pyrogen and endotoxins testing: questions and answers. https://www.fda.gov/downloads/drugs/guidances/ucm310098.pdf
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
Jones CF, Grainger DW (2009) In vitro assessments of nanomaterial toxicity. Adv Drug Deliv Rev 61(6):438–456. doi:10.1016/j.addr.2009.03.005
Sharma SK (1986) Endotoxin detection and elimination in biotechnology. Biotechnol Appl Biochem 8(1):5–22
Crist RM, Grossman JH, Patri AK, Stern ST, Dobrovolskaia MA, Adiseshaiah PP, Clogston JD, McNeil SE (2013) Common pitfalls in nanotechnology: lessons learned from NCI’s Nanotechnology Characterization Laboratory. Integr Biol (Camb) 5(1):66–73. doi:10.1039/c2ib20117h
Dobrovolskaia MA, Patri AK, Potter TM, Rodriguez JC, Hall JB, McNeil SE (2012) Dendrimer-induced leukocyte procoagulant activity depends on particle size and surface charge. Nanomedicine (Lond) 7(2):245–256. doi:10.2217/nnm.11.105
Inoue K (2011) Promoting effects of nanoparticles/materials on sensitive lung inflammatory diseases. Environ Health Prev Med 16(3):139–143. doi:10.1007/s12199-010-0177-7
Inoue K, Takano H (2011) Aggravating impact of nanoparticles on immune-mediated pulmonary inflammation. Scientific World J 11:382–390. doi:10.1100/tsw.2011.44
Inoue K, Takano H, Yanagisawa R, Hirano S, Kobayashi T, Fujitani Y, Shimada A, Yoshikawa T (2007) Effects of inhaled nanoparticles on acute lung injury induced by lipopolysaccharide in mice. Toxicology 238(2–3):99–110. doi:10.1016/j.tox.2007.05.022
Inoue K, Takano H, Yanagisawa R, Hirano S, Sakurai M, Shimada A, Yoshikawa T (2006) Effects of airway exposure to nanoparticles on lung inflammation induced by bacterial endotoxin in mice. Environ Health Perspect 114(9):1325–1330. doi:10.1289/ehp.8903
Shi Y, Yadav S, Wang F, Wang H (2010) Endotoxin promotes adverse effects of amorphous silica nanoparticles on lung epithelial cells in vitro. J Toxicol Environ Health A 73(11):748–756. doi:10.1080/15287391003614042
Dobrovolskaia MA, McNeil SE (2016) Nanoparticles and endotoxin. In: Dobrovolskaia MA, McNeil SE (eds) Handbook of immunological properties of engineered nanomaterials, vol 1. World Scientific Publishing, Singapore, pp 143–187
Alwis KU, Milton DK (2006) Recombinant factor C assay for measuring endotoxin in house dust: comparison with LAL, and (1 --> 3)-beta-D-glucans. Am J Ind Med 49(4):296–300. doi:10.1002/ajim.20264
Ding JL, Ho B (2010) Endotoxin detection--from limulus amebocyte lysate to recombinant factor C. Subcell Biochem 53:187–208. doi:10.1007/978-90-481-9078-2_9
McKenzie JH, Alwis KU, Sordillo JE, Kalluri KS, Milton DK (2011) Evaluation of lot-to-lot repeatability and effect of assay media choice in the recombinant Factor C assay. J Environ Monit 13(6):1739–1745. doi:10.1039/c1em10035a
Fujita Y, Nabetani T (2014) Iron sulfate inhibits Limulus activity by induction of structural and qualitative changes in lipid A. J Appl Microbiol 116(1):89–99. doi:10.1111/jam.12349
Reich J, Lang P, Grallert H, Motschmann H (2016) Masking of endotoxin in surfactant samples: effects on Limulus-based detection systems. Biologicals 44(5):417–422. doi:10.1016/j.biologicals.2016.04.012
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
Tran T, Beal SG (2016) Application of the 1,3-beta-D-Glucan (Fungitell) assay in the diagnosis of invasive fungal infections. Arch Pathol Lab Med 140(2):181–185. doi:10.5858/arpa.2014-0230-RS
Henne W, Schulze H, Pelger M, Tretzel J, von Sengbusch G (1984) Hollow-fiber dialyzers and their pyrogenicity testing by Limulus amebocyte lysate. Artif Organs 8(3):299–305
Neun BW, Dobrovolskaia MA (2011) Detection and quantitative evaluation of endotoxin contamination in nanoparticle formulations by LAL-based assays. Methods Mol Biol 697:121–130. doi:10.1007/978-1-60327-198-1_12
Sandle T (2011) A practical approach to depyrogenation studies using bacterial endotoxin. J GxP Compliance 15(4):90–96
Subbarao N (2016) Nanoparticle sterility and sterilization of nanomaterials. In: Dobrovolskaia MA, McNeil SE (eds) Handbook of immunological properties of engineered nanomaterials, vol 1 and 6. World Scientific Publishing Ltd, Singapore, pp 53–75
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
Ragab AA, Van De Motter R, Lavish SA, Goldberg VM, Ninomiya JT, Carlin CR, Greenfield EM (1999) Measurement and removal of adherent endotoxin from titanium particles and implant surfaces. J Orthop Res 17(6):803–809. doi:10.1002/jor.1100170603
Dobrovolskaia MA, Neun BW, Clogston JD, Ding H, Ljubimova J, McNeil SE (2010) Ambiguities in applying traditional Limulus amebocyte lysate tests to quantify endotoxin in nanoparticle formulations. Nanomedicine (Lond) 5(4):555–562. doi:10.2217/nnm.10.29
London AS, Mackay K, Lihon M, He Y, Alabi BR (2014) Gel filtration chromatography as a method for removing bacterial endotoxin from antibody preparations. Biotechnol Prog 30(6):1497–1501. doi:10.1002/btpr.1961
Ma R, Zhao J, Du HC, Tian S, Li LW (2012) Removing endotoxin from plasmid samples by Triton X-114 isothermal extraction. Anal Biochem 424(2):124–126. doi:10.1016/j.ab.2012.02.015
Mack L, Brill B, Delis N, Groner B (2014) Endotoxin depletion of recombinant protein preparations through their preferential binding to histidine tags. Anal Biochem 466:83–88. doi:10.1016/j.ab.2014.08.020
Magalhaes PO, Lopes AM, Mazzola PG, Rangel-Yagui C, Penna TC, Pessoa A Jr (2007) Methods of endotoxin removal from biological preparations: a review. J Pharm Pharm Sci 10(3):388–404
Afonin KA, Grabow WW, Walker FM, Bindewald E, Dobrovolskaia MA, Shapiro BA, Jaeger L (2011) Design and self-assembly of siRNA-functionalized RNA nanoparticles for use in automated nanomedicine. Nat Protoc 6(12):2022–2034. doi:10.1038/nprot.2011.418
Dobrovolskaia MA, Germolec DR, Weaver JL (2009) Evaluation of nanoparticle immunotoxicity. Nat Nanotechnol 4(7):411–414. doi:10.1038/nnano.2009.175
Acknowledgment
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Neun, B.W., Dobrovolskaia, M.A. (2018). Considerations and Some Practical Solutions to Overcome Nanoparticle Interference with LAL Assays and to Avoid Endotoxin Contamination in Nanoformulations. In: McNeil, S. (eds) Characterization of Nanoparticles Intended for Drug Delivery. Methods in Molecular Biology, vol 1682. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7352-1_3
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7352-1_3
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7350-7
Online ISBN: 978-1-4939-7352-1
eBook Packages: Springer Protocols