Modular adjuvant-free pan-HLA-DR-immunotargeting subunit vaccine against SARS-CoV-2 elicits broad sarbecovirus-neutralizing antibody responses

Kassardjian, Audrey; Sun, Eric; Sookhoo, Jamie; Muthuraman, Krithika; Boligan, Kayluz Frias; Kucharska, Iga; Rujas, Edurne; Jetha, Arif; Branch, Donald R.; Babiuk, Shawn; Barber, Brian H.; Julien, Jean-Philippe

doi:10.1016/j.celrep.2023.112391

Cited by 7 publications

(5 citation statements)

References 80 publications

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“…We previously used mAb 44H10 as the scaffold of an immunotargeting vaccine (ITV) against SARS-CoV-2, whereby the SARS-CoV-2 Spike protein RBD was fused to a chimeric mouse-human 44H10 IgG, effectively targeting this antigen to MHC class II-expressing antigen-presenting cells [ 23 ]. Here, the humanization of the 44H10 antibody benefits the development of the ITV platform for potential use in humans by minimizing the risk of immunogenicity of the antibody scaffold and focusing the response on the immunotargeted cargo antigen.…”

Section: Discussionmentioning

confidence: 99%

“…HLA-DR α and β chain plasmids (previously described [ 23 ]) were co-transfected in a 1:1 ratio (50 µg total per 200 mL of cell culture) into HEK 293S (GnT I −/− ) cells. Recombinant HLA-DR was purified by affinity chromatography via a HisTrap Ni-NTA column (Cytiva) and eluted using 20 mM Tris, pH 8.0, 500 mM imidazole.…”

Section: Methodsmentioning

confidence: 99%

“…The antibody of interest in this report, mAb 44H10, was isolated from BALB/c mice immunized with cells from a non-T, non-B acute lymphoblastic leukemia (ALL) cell line and further discovered to react to MHC class II molecules by radioimmunoprecipitation and tissue distribution studies [ 22 ]. Murine mAb 44H10 was shown to target an epitope on the HLA-DR molecule conserved in humans [ 23 , 24 ], suggesting its potential clinical benefit across populations irrespective of HLA haplotype. Previously, this mAb was used as the scaffold for immunotargeting vaccines delivering protein antigens to MHC class II-expressing antigen-presenting cells [ 23 , 25 ], successfully inducing potent adjuvant-free antigen-specific antibody responses.…”

Section: Introductionmentioning

confidence: 99%

“…Murine mAb 44H10 was shown to target an epitope on the HLA-DR molecule conserved in humans [ 23 , 24 ], suggesting its potential clinical benefit across populations irrespective of HLA haplotype. Previously, this mAb was used as the scaffold for immunotargeting vaccines delivering protein antigens to MHC class II-expressing antigen-presenting cells [ 23 , 25 ], successfully inducing potent adjuvant-free antigen-specific antibody responses. As a highly modular platform with several cargo anchoring points, mAb 44H10 could be employed for the delivery of diverse antigens, with potential applications in the development of both vaccines and therapeutics.…”

Section: Introductionmentioning

confidence: 99%

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Humanization of Pan-HLA-DR mAb 44H10 Hinges on Critical Residues in the Antibody Framework

Kassardjian,

Ivanochko,

Barber

et al. 2024

Antibodies

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Reducing the immunogenicity of animal-derived monoclonal antibodies (mAbs) for use in humans is critical to maximize therapeutic effectiveness and preclude potential adverse events. While traditional humanization methods have primarily focused on grafting antibody Complementarity-Determining Regions (CDRs) on homologous human antibody scaffolds, framework regions can also play essential roles in antigen binding. Here, we describe the humanization of the pan-HLA-DR mAb 44H10, a murine antibody displaying significant involvement of the framework region in antigen binding. Using a structure-guided approach, we identify and restore framework residues that directly interact with the antigen or indirectly modulate antigen binding by shaping the antibody paratope and engineer a humanized antibody with affinity, biophysical profile, and molecular binding basis comparable to that of the parental 44H10 mAb. As a humanized molecule, this antibody holds promise as a scaffold for the development of MHC class II-targeting therapeutics and vaccines.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Humanization of Pan-HLA-DR mAb 44H10 Hinges on Critical Residues in the Antibody Framework

Kassardjian,

Ivanochko,

Barber

et al. 2024

Antibodies

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“…S17). The c44H10 Fab was co-purified with the TRACeR-MHC complex to facilitate particle alignment and achieve high resolution 39 . As expected, the ternary complex structure reveals that the binding mode of 𝑇𝑅𝐴𝐶𝑒𝑅 !…”

Section: Structural Validation By Cryo-electron Microscopymentioning

confidence: 99%

A general platform for targeting MHC-II antigens via a single loop

Du,

Liu,

Jude

et al. 2024

Preprint

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Class-II major histocompatibility complexes (MHC-IIs) are central to the communications between CD4+ T cells and antigen presenting cells (APCs), but intrinsic structural features associated with MHC-II make it difficult to develop a general targeting system with high affinity and antigen specificity. Here, we introduce a protein platform, Targeted Recognition of Antigen-MHC Complex Reporter for MHC-II (TRACeR-II), to enable the rapid development of peptide-specific MHC-II binders. TRACeR-II has a small helical bundle scaffold and uses an unconventional mechanism to recognize antigens via a single loop. This unique antigen-recognition mechanism renders this platform highly versatile and amenable to direct structural modeling of the interactions with the antigen. We demonstrate that TRACeR-II binders can be rapidly evolved across multiple alleles, while computational protein design can produce specific binding sequences for a SARS-CoV-2 peptide of unknown complex structure. TRACeR-II sheds light on a simple and straightforward approach to address the MHC peptide targeting challenge, without relying on combinatorial selection on complementarity determining region (CDR) loops. It presents a promising basis for further exploration in immune response modulation as well as a broad range of theragnostic applications.

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