Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis
Developing a preventative vaccine for HIV-1 has been a global priority. The elicitation of broadly neutralizing antibodies (bNAbs) against a broad range of HIV-1 envelopes (Envs) from various strains appears to be a critical requirement for an efficacious HIV-1 vaccine. To understand their ability t...
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| Format: | Article |
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MDPI AG
2023-12-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/13/1/33 |
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| author | Shraddha Basu Neelakshi Gohain Jiae Kim Hung V. Trinh Misook Choe M. Gordon Joyce Mangala Rao |
| author_facet | Shraddha Basu Neelakshi Gohain Jiae Kim Hung V. Trinh Misook Choe M. Gordon Joyce Mangala Rao |
| author_sort | Shraddha Basu |
| collection | DOAJ |
| description | Developing a preventative vaccine for HIV-1 has been a global priority. The elicitation of broadly neutralizing antibodies (bNAbs) against a broad range of HIV-1 envelopes (Envs) from various strains appears to be a critical requirement for an efficacious HIV-1 vaccine. To understand their ability to neutralize HIV-1, it is important to characterize the binding characteristics of bNAbs. Our work is the first to utilize microscale thermophoresis (MST), a rapid, economical, and flexible in-solution temperature gradient method to quantitatively determine the binding affinities of bNAbs and non-neutralizing monoclonal antibodies (mAbs) to HIV-1 recombinant envelope monomer and trimer proteins of different subtypes, pseudoviruses (PVs), infectious molecular clones (IMCs), and cells expressing the trimer. Our results demonstrate that the binding affinities were subtype-dependent. The bNAbs exhibited a higher affinity to IMCs compared to PVs and recombinant proteins. The bNAbs and mAbs bound with high affinity to native-like gp160 trimers expressed on the surface of CEM cells compared to soluble recombinant proteins. Interesting differences were seen with V2-specific mAbs. Although they recognize linear epitopes, one of the antibodies also bound to the Envs on PVs, IMCs, and a recombinant trimer protein, suggesting that the epitope was not occluded. The identification of epitopes on the envelope surface that can bind to high affinity mAbs could be useful for designing HIV-1 vaccines and for down-selecting vaccine candidates that can induce high affinity antibodies to the HIV-1 envelope in their native conformation. |
| first_indexed | 2024-03-08T15:10:33Z |
| format | Article |
| id | doaj.art-94b557cde4d44f99a6f099511f0e68ac |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-08T15:10:33Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-94b557cde4d44f99a6f099511f0e68ac2024-01-10T14:53:15ZengMDPI AGCells2073-44092023-12-011313310.3390/cells13010033Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale ThermophoresisShraddha Basu0Neelakshi Gohain1Jiae Kim2Hung V. Trinh3Misook Choe4M. Gordon Joyce5Mangala Rao6Henry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 20817, USAHenry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 20817, USAHenry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 20817, USAHenry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 20817, USAHenry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 20817, USAHenry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 20817, USAUS Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USADeveloping a preventative vaccine for HIV-1 has been a global priority. The elicitation of broadly neutralizing antibodies (bNAbs) against a broad range of HIV-1 envelopes (Envs) from various strains appears to be a critical requirement for an efficacious HIV-1 vaccine. To understand their ability to neutralize HIV-1, it is important to characterize the binding characteristics of bNAbs. Our work is the first to utilize microscale thermophoresis (MST), a rapid, economical, and flexible in-solution temperature gradient method to quantitatively determine the binding affinities of bNAbs and non-neutralizing monoclonal antibodies (mAbs) to HIV-1 recombinant envelope monomer and trimer proteins of different subtypes, pseudoviruses (PVs), infectious molecular clones (IMCs), and cells expressing the trimer. Our results demonstrate that the binding affinities were subtype-dependent. The bNAbs exhibited a higher affinity to IMCs compared to PVs and recombinant proteins. The bNAbs and mAbs bound with high affinity to native-like gp160 trimers expressed on the surface of CEM cells compared to soluble recombinant proteins. Interesting differences were seen with V2-specific mAbs. Although they recognize linear epitopes, one of the antibodies also bound to the Envs on PVs, IMCs, and a recombinant trimer protein, suggesting that the epitope was not occluded. The identification of epitopes on the envelope surface that can bind to high affinity mAbs could be useful for designing HIV-1 vaccines and for down-selecting vaccine candidates that can induce high affinity antibodies to the HIV-1 envelope in their native conformation.https://www.mdpi.com/2073-4409/13/1/33HIV-1 epitopesMSTbinding affinityrecombinant HIV-1 proteinspseudovirusinfectious molecular clone |
| spellingShingle | Shraddha Basu Neelakshi Gohain Jiae Kim Hung V. Trinh Misook Choe M. Gordon Joyce Mangala Rao Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis Cells HIV-1 epitopes MST binding affinity recombinant HIV-1 proteins pseudovirus infectious molecular clone |
| title | Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis |
| title_full | Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis |
| title_fullStr | Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis |
| title_full_unstemmed | Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis |
| title_short | Determination of Binding Affinity of Antibodies to HIV-1 Recombinant Envelope Glycoproteins, Pseudoviruses, Infectious Molecular Clones, and Cell-Expressed Trimeric gp160 Using Microscale Thermophoresis |
| title_sort | determination of binding affinity of antibodies to hiv 1 recombinant envelope glycoproteins pseudoviruses infectious molecular clones and cell expressed trimeric gp160 using microscale thermophoresis |
| topic | HIV-1 epitopes MST binding affinity recombinant HIV-1 proteins pseudovirus infectious molecular clone |
| url | https://www.mdpi.com/2073-4409/13/1/33 |
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