Design of the SARS-CoV-2 RBD vaccine antigen improves neutralizing antibody response

Dickey, Thayne H.; Tang, Wai Kwan; Butler, Brandi; Ouahes, Tarik; Orr-Gonzalez, Sachy; Salinas, Nichole D.; Lambert, Lynn; Tolia, Niraj H.

doi:10.1126/sciadv.abq8276

Cited by 37 publications

(39 citation statements)

References 83 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Griffoni et al have reported that T cells can recognize up to 10 different SARS-CoV-2 proteins, with spike being the most immunodominant, followed by nucleocapsid, membrane, and other non-structural proteins (149,150). These data, together with the fact that the receptor binding domain (RBD) of the SARS-CoV-2 spike protein is the main target of neutralizing antibodies (53,151) (153)(154)(155)(156)(157)(158). Since vaccination protocols started in December 2020 with BNT162b2 vaccine, it has been estimated that global anti-SARS-CoV-2 vaccination saved nearly 20 million lives in its first year of application (159).…”

Section: T Cell Response After Vaccinationmentioning

confidence: 99%

Defending against SARS-CoV-2: The T cell perspective

2023

View full text Add to dashboard Cite

SARS-CoV-2-specific T cell response has been proven essential for viral clearance, COVID-19 outcome and long-term memory. Impaired early T cell-driven immunity leads to a severe form of the disease associated with lymphopenia, hyperinflammation and imbalanced humoral response. Analyses of acute SARS-CoV-2 infection have revealed that mild COVID-19 course is characterized by an early induction of specific T cells within the first 7 days of symptoms, coordinately followed by antibody production for an effective control of viral infection. In contrast, patients who do not develop an early specific cellular response and initiate a humoral immune response with subsequent production of high levels of antibodies, develop severe symptoms. Yet, delayed and persistent bystander CD8+ T cell activation has been also reported in hospitalized patients and could be a driver of lung pathology. Literature supports that long-term maintenance of T cell response appears more stable than antibody titters. Up to date, virus-specific T cell memory has been detected 22 months post-symptom onset, with a predominant IL-2 memory response compared to IFN-γ. Furthermore, T cell responses are conserved against the emerging variants of concern (VoCs) while these variants are mostly able to evade humoral responses. This could be partly explained by the high HLA polymorphism whereby the viral epitope repertoire recognized could differ among individuals, greatly decreasing the likelihood of immune escape. Current COVID-19-vaccination has been shown to elicit Th1-driven spike-specific T cell response, as does natural infection, which provides substantial protection against severe COVID-19 and death. In addition, mucosal vaccination has been reported to induce strong adaptive responses both locally and systemically and to protect against VoCs in animal models. The optimization of vaccine formulations by including a variety of viral regions, innovative adjuvants or diverse administration routes could result in a desirable enhanced cellular response and memory, and help to prevent breakthrough infections. In summary, the increasing evidence highlights the relevance of monitoring SARS-CoV-2-specific cellular immune response, and not only antibody levels, as a correlate for protection after infection and/or vaccination. Moreover, it may help to better identify target populations that could benefit most from booster doses and to personalize vaccination strategies.

show abstract

Section: T Cell Response After Vaccinationmentioning

confidence: 99%

Defending against SARS-CoV-2: The T cell perspective

2023

View full text Add to dashboard Cite

show abstract

“…This protein is considered an important target for developing a SARS subunit vaccine. Indeed, the majority of the potent neutralising antibodies against SARS-CoV-2 target the RBD 30 , but reports differ on whether RBD alone or full S1 subunits of the spike are more potent vaccines in terms of IgG and IgA induction 31,32 .To explore this concept, we expressed both RBD and this S1 subunit from the pALiCE02 vector and with different HaloTag and Strep-II tag conformations. Antigen reactivity to two commercial antibodies was initially assessed, selected based on the recognition of either linear or conformational RBD epitopes.…”

Section: N-linked Glycosylation Of Glycoproteins Expressed In Bylmentioning

confidence: 99%

ALiCE^®: A versatile, high yielding and scalable eukaryotic cell-free protein synthesis (CFPS) system

Gupta¹,

Flaskamp²,

Roentgen³

et al. 2022

Preprint

View full text Add to dashboard Cite

Eukaryotic cell-free protein synthesis (CFPS) systems have the potential to simplify and speed up the expression and high-throughput analysis of complex proteins with functionally relevant post-translational modifications (PTMs). However, low yields and the inability to scale such systems have so far prevented their widespread adoption in protein research and manufacturing. Here, we present a detailed demonstration for the capabilities of a CFPS system derived from Nicotiana tabacum BY-2 cell culture (BY-2 lysate; BYL). BYL is able to express diverse, functional proteins at high yields in under 48 hours, complete with native disulfide bonds and N-glycosylation. An optimised version of the technology is commercialised as 'ALiCE', engineered for high yields of up to 3 mg/mL. Recent advances in the scaling of BYL production methodologies have allowed scaling of the CFPS reaction. We show simple, linear scale-up of batch mode reporter proten expression from a 100 uL microtiter plate format to 10 mL and 100 mL volumes in standard Erlenmeyer flasks, culminating in preliminary data from 1 L reactions in a CELL-tainer CT20 rocking motion bioreactor. As such, these works represent the first published example of a eukaryotic CFPS reaction scaled past the 10 mL level by several orders of magnitude. We show the ability of BYL to produce the simple reporter protein eYFP and large, multimeric virus-like particles directly in the cytosolic fraction. Complex proteins are processed using the native microsomes of BYL and functional expression of multiple classes of complex, difficult-to-express proteins is demonstrated, specifically: a dimeric, glycoprotein enzyme, glucose oxidase; the monoclonal antibody adalimumab; the SARS-Cov-2 receptor-binding domain; human epidermal growth factor; and a G protein-coupled receptor membrane protein, cannabinoid receptor type 2. Functional binding and activity are shown using a combination of surface plasmon resonance techniques, a serology-based ELISA method and a G protein activation assay. Finally, in-depth post-translational modification (PTM) characterisation of purified proteins through disulfide bond and N-glycan analysis is also revealed - previously difficult in the eukaryotic CFPS space due to limitations in reaction volumes and yields. Taken together, BYL provides a real opportunity for screening of complex proteins at the microscale with subsequent amplification to manufacturing-ready levels using off-the-shelf protocols. This end-to-end platform suggests the potential to significantly reduce cost and the time-to-market for high value proteins and biologics.

show abstract

“…Stabilizer for Protein Expression and Epitope Design (SPEEDesign) is a ROSETTA-based computational and in vitro screening pipeline that designs antigens resulting in potent and durable vaccines 16 . The objectives of SPEEDesign are to improve the protective efficacy of a given antigen by: (1) focusing the immune response towards potently neutralizing antibody epitopes on the antigen; (2) reducing or eliminating immune responses to poorly neutralizing and/or immunodominant epitopes within the antigen; (3) optimizing the thermal stability of the antigen to increase its durability in vivo following immunization; (4) promoting conformational states of a protein antigen that may be hidden, for example by removing the glycan on pfs48/45 D3 that may mask neutralizing epitopes.…”

Section: Introductionmentioning

confidence: 99%

“…The objectives of SPEEDesign are to improve the protective efficacy of a given antigen by: (1) focusing the immune response towards potently neutralizing antibody epitopes on the antigen; (2) reducing or eliminating immune responses to poorly neutralizing and/or immunodominant epitopes within the antigen; (3) optimizing the thermal stability of the antigen to increase its durability in vivo following immunization; (4) promoting conformational states of a protein antigen that may be hidden, for example by removing the glycan on pfs48/45 D3 that may mask neutralizing epitopes. SPEEDesign has proven successful in the development of improved SARS-CoV-2 antigens 16 .…”

Section: Introductionmentioning

confidence: 99%

Design of a stabilized non-glycosylated Pfs48/45 antigen enables a potent malaria transmission-blocking nanoparticle vaccine

et al. 2023

Self Cite

View full text Add to dashboard Cite

A malaria vaccine that blocks parasite transmission from human to mosquito would be a powerful method of disrupting the parasite lifecycle and reducing the incidence of disease in humans. Pfs48/45 is a promising antigen in development as a transmission blocking vaccine (TBV) against the deadliest malaria parasite Plasmodium falciparum. The third domain of Pfs48/45 (D3) is an established TBV candidate, but production challenges have hampered development. For example, to date, a non-native N-glycan is required to stabilize the domain when produced in eukaryotic systems. Here, we implement a SPEEDesign computational design and in vitro screening pipeline that retains the potent transmission blocking epitope in Pfs48/45 while creating a stabilized non-glycosylated Pfs48/45 D3 antigen with improved characteristics for vaccine manufacture. This antigen can be genetically fused to a self-assembling single-component nanoparticle, resulting in a vaccine that elicits potent transmission-reducing activity in rodents at low doses. The enhanced Pfs48/45 antigen enables many new and powerful approaches to TBV development, and this antigen design method can be broadly applied towards the design of other vaccine antigens and therapeutics without interfering glycans.

show abstract

Design of the SARS-CoV-2 RBD vaccine antigen improves neutralizing antibody response

Cited by 37 publications

References 83 publications

Defending against SARS-CoV-2: The T cell perspective

Defending against SARS-CoV-2: The T cell perspective

ALiCE^®: A versatile, high yielding and scalable eukaryotic cell-free protein synthesis (CFPS) system

Design of a stabilized non-glycosylated Pfs48/45 antigen enables a potent malaria transmission-blocking nanoparticle vaccine

Contact Info

Product

Resources

About

Design of the SARS-CoV-2 RBD vaccine antigen improves neutralizing antibody response

Cited by 37 publications

References 83 publications

Defending against SARS-CoV-2: The T cell perspective

Defending against SARS-CoV-2: The T cell perspective

ALiCE®: A versatile, high yielding and scalable eukaryotic cell-free protein synthesis (CFPS) system

Design of a stabilized non-glycosylated Pfs48/45 antigen enables a potent malaria transmission-blocking nanoparticle vaccine

Contact Info

Product

Resources

About

ALiCE^®: A versatile, high yielding and scalable eukaryotic cell-free protein synthesis (CFPS) system