Abstract
Drug delivery is of paramount importance, since the drug needs to be delivered to a specific site, in adequate concentration, avoiding degradation in order to provide therapeutic efficacy. Different nanocarriers have been used over the years for this purpose and liposomes are well-established systems due to the high biocompatibility and the possibility to vehiculate both hydrophilic and lipophilic drugs. In order to circumvent the rapid clearance by the reticuloendothelial system and to avoid the healthy cells exposure to the drug, long circulating liposomes containing polyethyleneglycol (PEG) and functionalized liposomes for targeted delivery have been developed. Immunoliposomes consist of liposomes containing antibodies or antibody fragments attached at the membrane surface. This attachment can be performed using PEG lipids, containing a reactive terminal group such as maleimide and thiolated antibodies. Additionaly, the use of PEG chains as spacers increases antibody–antigen affinity, since the antibody is not shielded by the steric hindrance of PEG and also due to the correct orientation of antibodies for interaction with receptors on cell surface. In this chapter, we describe and discuss in details the protocol to prepare anti-epidermal growth factor receptor (anti-EGFR) and anti-human epidermal growth factor receptor 2 (anti-HER2) liposomes using cetuximab and trastuzumab as antibodies. We present the direct coupling method based on the maleimide thioether reaction for these immunoliposomes preparation and present some characterization steps and in vitro studies in cell culture which can be used for better understanding these nanocarriers.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Fomina N, Sankaranarayanan J, Almutairi A (2012) Photochemical mechanisms of light-triggered release from nanocarriers. Adv Drug Deliv Rev 64(11):1005–1020
Eloy JO, Claro de Souza M, Petrilli R, Barcellos JPA, Lee RJ, Marchetti JM (2014) Liposomes as carriers of hydrophilic small molecule drugs: strategies to enhance encapsulation and delivery. Colloids Surf B Biointerfaces 123:345–363
Torchilin VP (2005) Recent advances with liposomes as pharmaceutical carriers. Nat Rev Drug Discov 4(2):145–160
Chang HI, Yeh MK (2012) Clinical development of liposome-based drugs: formulation, characterization, and therapeutic efficacy. Int J Nanomedicine 7:49–60
Kwon SS, Kim SY, Kong BJ, Jin K, Noh GY, Im NR, Lim JW, Ha JH, Kim J, Park SN (2015) Cell-penetrating peptide-conjugated liposomes as transdermal delivery system of Polygonum aviculare L. extract. Int J Pharm 483:26–37
Mendonça LS, Firmino F, Moreira JN, De Lima MCP (2010) Transferrin receptor-targeted liposomes encapsulating anti- BCR-ABL siRNA or asODN for chronic myeloid leukemia treatment. Bioconjug Chem 21:157–168
Li H, Piao L, Yu B, Yung BC, Zhang W, Wang PG, Lee JL, Lee RJ (2011) Delivery of calf thymus DNA to tumor by folate receptor targeted cationic liposomes. Biomaterials 32(27):6614–6620
Wijagkanalan W, Kawakami S, Higuchi Y, Yamashita F, Hashida M (2011) Intratracheally instilled mannosylated cationic liposome/NF-κB decoy complexes for effective prevention of LPS-induced lung inflammation. J Control Release 149(1):42–50
Loureiro JA, Gomes B, Fricker G, Cardoso I, Ribeiro CA, Gaiteiro C, Coelho MA, Pereira MC, Rocha S (2015) Dual ligand immunoliposomes for drug delivery to the brain. Colloids Surf B Biointerfaces 134:213–219
Petrilli R, Eloy JO, Marchetti JM, Lopez RFV, Lee RJ (2014) Targeted lipid nanoparticles for antisense oligonucleotide delivery. Curr Pharm Biotechnol 15(9):847–855
Kirpotin DB, Drummond DC, Shao Y, Shalaby MR, Hong K, Nielsen UB, Marks JD, Benz CC, Park JW (2006) Antibody targeting of long-circulating lipidic nanoparticles does not increase tumor localization but does increase internalization in animal models. Cancer Res 66(13):6732–6740
Pan X, Wu G, Yang W, Barth RF, Tjarks W, Lee J (2007) Synthesis of cetuximab-immunoliposomes via a cholesterol- based membrane anchor for targeted delivery of a neutron capture therapy (NCT) agent to glioma cells. Bioconjug Chem 18(1):101–108
Demarest SJ, Hariharan K, Dong J (2011) Emerging antibody combinations in oncology. MAbs 3(4):338–351
Petrilli R, Eloy J, Lopez RFV, Lee R (2017) Cetuximab immunoliposomes enhance delivery of 5-FU to skin squamous carcinoma cells. Anti Cancer Agents Med Chem 17:301–308
Eloy J, Petrilli R, Brueggemeier RW, Marchetti, JM, Lee RJ (2017) Rapamycin loaded immunoliposomes functionalized with trastuzumab: a strategy to enhance cytotoxicity to HER2 positive breast cancer cells. Anti Cancer Agents Med Chem 17:48–56
Chang HR (2010) Trastuzumab-based neoadjuvant therapy in patients with HER2-positive breast cancer. Cancer 116(12):2856–2867
Cohen MH, Chen H, Shord S, Fuchs C, He K, Zhao H, Sickafuse S, Keegan P, Pazdur R (2013) Approval summary: Cetuximab in combination with cisplatin or carboplatin and 5-fluorouracil for the first-line treatment of patients with recurrent locoregional or metastatic squamous cell head and neck cancer. Oncologist 18:460–466
Schnyder A, Krähenbühl S, Török M, Drewe J, Huwyler J (2004) Targeting of skeletal muscle in vitro using biotinylated immunoliposomes. Biochem J 377:61–67
Koning GA, Kamps JAAM, Scherphof GL (2002) Efficient intracellular delivery of 5-fluorodeoxyuridine into colon cancer cells by targeted immunoliposomes. Cancer Detect Prev 26(4):299–307
Nobs L, Buchegger F, Gurny R, Alle E (2004) Current methods for attaching targeting ligands to liposomes and nanoparticles. J Pharm Sci 93(8):1980–1992
Acknowledgments
The authors would like to acknowledge grants #2012/23764-3, #2012/10388-3, #2012/21513-3, #2013/15134-2, and #2014/22451-7 from Sao Paulo Research Foundation (FAPESP) and CNPQ for grant #480962/2013-8.
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
Petrilli, R., Eloy, J.O., Lee, R.J., Lopez, R.F.V. (2018). Preparation of Immunoliposomes by Direct Coupling of Antibodies Based on a Thioether Bond. In: Picanço-Castro, V., Swiech, K. (eds) Recombinant Glycoprotein Production. Methods in Molecular Biology, vol 1674. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7312-5_19
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7312-5_19
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7311-8
Online ISBN: 978-1-4939-7312-5
eBook Packages: Springer Protocols