Conjugation of native‐like hiv‐1 envelope trimers onto liposomes using edc/sulfo‐nhs chemistry: Requirements and limitations

Suleiman E, Mayer J, Lehner E, Kohlhauser B, Katholnig A, Batzoni M, Damm D, Temchura V, Wagner A, Überla K, Vorauer‐uhl K (2020)

Publication Type: Journal article

Publication year: 2020

Journal

Pharmaceutics MDPI

Book Volume: 12

Pages Range: 1-27

Article Number: 979

Journal Issue: 10

DOI: 10.3390/pharmaceutics12100979

Abstract

The display of native‐like human immunodeficiency virus type 1 envelope (HIV‐1 Env) trimers on liposomes has gained wide attention over the last few years. Currently, available methods have enabled the preparation of Env‐liposome conjugates of unprecedented quality. However, these protocols require the Env trimer to be tagged and/or to carry a specific functional group. For this reason, we have investigated N‐(3‐Dimethylaminopropyl)‐N′‐ethylcarbodiimide/N‐ Hydroxysulfosuccinimide (EDC/Sulfo‐NHS) chemistry for its potential to covalently conjugate tag-free, non‐functionalized native‐like Env trimers onto the surface of carboxyl‐functionalized liposomes. The preservation of the liposome’s physical integrity and the immunogen’s conformation required a fine‐tuned two‐step approach based on the controlled use of β‐ mercaptoethanol. The display of Env trimers was strictly limited to activated liposomes of positive charge, i.e., liposomes with a positive zeta potential that carry amine‐reactive Sulfo‐NHS esters on their surface. In agreement with that, conjugation was found to be highly ionic strength‐ and pH-dependent. Overall, we have identified electrostatic pre‐concentration (i.e., close proximity between negatively charged Env trimers and positively charged liposomes established through electrostatic attraction) to be crucial for conjugation reactions to proceed. The present study highlights the requirements and limitations of potentially scalable EDC/Sulfo‐NHS‐based approaches and represents a solid basis for further research into the controlled conjugation of tag‐free, non-functionalized native‐like Env trimers on the surface of liposomes, and other nanoparticles.

Authors with CRIS profile

Dominik Damm Institute of Clinical and Molecular Virology Vladimir Temchura Institute of Clinical and Molecular Virology Klaus Überla Lehrstuhl für Klinische und Molekulare Virologie

Involved external institutions

Polymun Scientific Immunbiologische Forschung GmbH AT Austria (AT) Universität für Bodenkultur Wien (BOKU) / University of Natural Resources and Life Sciences, Vienna AT Austria (AT)

How to cite

APA:

Suleiman, E., Mayer, J., Lehner, E., Kohlhauser, B., Katholnig, A., Batzoni, M.,... Vorauer‐uhl, K. (2020). Conjugation of native‐like hiv‐1 envelope trimers onto liposomes using edc/sulfo‐nhs chemistry: Requirements and limitations. Pharmaceutics, 12(10), 1-27. https://doi.org/10.3390/pharmaceutics12100979

MLA:

Suleiman, Ehsan, et al. "Conjugation of native‐like hiv‐1 envelope trimers onto liposomes using edc/sulfo‐nhs chemistry: Requirements and limitations." Pharmaceutics 12.10 (2020): 1-27.

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