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Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay
Published on: September 14, 2014
Screening the interactions between HIV-1 neutralizing antibodies and model lipid surfaces
Gregory J Hardy1, Yee Lam, Shelley M Stewart
1Mechanical Engineering and Materials Science, Duke University, Durham, NC, USA.
Journal of Immunological Methods
|October 29, 2011
Summary
Developing broadly neutralizing antibodies (NAbs) for HIV-1 is challenging due to their lipid interactions. This study introduces a new assay to screen antibodies for effective lipid binding, aiding NAb development.
Area of Science:
- Immunology and Virology
- Biophysical Chemistry
- Antibody Engineering
Background:
- Broadly neutralizing antibodies (NAbs) targeting HIV-1, such as 4E10 and 2F5, interact with viral lipid membranes as a crucial initial step for neutralization.
- The development of such NAbs may be hindered by immunologic tolerance, potentially caused by autoreactivity with host cell membranes.
- Limited experimental data exists on the specific interactions between NAbs and lipid membranes, despite their significance.
Purpose of the Study:
- To develop a simple and efficient screening assay for identifying antibodies with lipid reactivity similar to known broadly neutralizing antibodies (NAbs).
- To investigate the role of charge and hydrophobicity in antibody-surface interactions relevant to NAb binding to lipid membranes.
- To provide a tool for screening interactions of monoclonal antibodies (mAbs) and lipid-reactive NAbs.
Main Methods:
- Development of a surface plasmon resonance (SPR) spectroscopy-based assay.
- Utilizing thiol self-assembled monolayers (SAMs) to mimic the lipid surface chemistries of HIV-1 membranes.
- Probing antibody binding to these SAMs to assess the influence of surface charge and hydrophobicity.
Main Results:
- NAb binding to hydrophobic thiol surfaces was significantly higher compared to control monoclonal antibodies (mAbs).
- The assay confirmed the importance of charge-mediated interactions in antibody-surface binding, consistent with previous findings on lipid vesicles.
- The developed SAMs effectively replicate the binding behavior of NAbs on lipid surfaces.
Conclusions:
- The SPR-based assay using thiol self-assembled monolayers (SAMs) is an efficient tool for screening antibody-lipid interactions.
- This method facilitates the selection of antibodies with specific lipid-binding properties relevant to HIV-1 neutralization.
- The findings highlight the critical role of lipid interactions in NAb function and provide a pathway for improved NAb development.
