Pharmaceutical Chemistry Scholarly Workshttps://hdl.handle.net/1808/812024-03-29T14:38:49Z2024-03-29T14:38:49ZEffects of aluminum-salt, CpG and emulsion adjuvants on the stability and immunogenicity of a virus-like particle displaying the SARS-CoV-2 receptor-binding domain (RBD) (Dataset)Kumru, Ozan S.Bajoria, SakshiKaur, KawaljitHickey, John M.Van Slyke, GretaDoering, JenniferBerman, KatherineRichardson, CharlesLien, HansKleanthous, HarryMantis, Nicholas J.Joshi, Sangeeta B.Volkin, David B.https://hdl.handle.net/1808/347952023-10-12T19:46:55Z2023-10-01T00:00:00ZEffects of aluminum-salt, CpG and emulsion adjuvants on the stability and immunogenicity of a virus-like particle displaying the SARS-CoV-2 receptor-binding domain (RBD) (Dataset)
Kumru, Ozan S.; Bajoria, Sakshi; Kaur, Kawaljit; Hickey, John M.; Van Slyke, Greta; Doering, Jennifer; Berman, Katherine; Richardson, Charles; Lien, Hans; Kleanthous, Harry; Mantis, Nicholas J.; Joshi, Sangeeta B.; Volkin, David B.
Second-generation COVID-19 vaccines with improved immunogenicity (e.g., breadth, duration) and availability (e.g., lower costs, refrigerator stable) are needed to enhance global coverage. In this work, we formulated a clinical-stage SARS-CoV-2 receptor binding domain (RBD) virus-like particle (VLP) vaccine candidate (IVX-411) with widely available adjuvants. Specifically, we assessed the in vitro storage stability and in vivo mouse immunogenicity of IVX-411 formulated with aluminum-salt adjuvants (Alhydrogel™, AH and Adjuphos™, AP), without or with the TLR-9 agonist CpG-1018™ (CpG), and compared these profiles to IVX-411 adjuvanted with an oil-in-water nano-emulsion (AddaVax™, AV). Although IVX-411 bound both AH and AP, lower binding strength of antigen to AP was observed by Langmuir binding isotherms. Interestingly, AH- and AP-adsorbed IVX-411 had similar storage stability profiles as measured by antigen binding assays (competitive ELISAs), but the latter displayed higher pseudovirus neutralizing titers (pNT) in mice, at levels comparable to titers elicited by AV-adjuvanted IVX-411. CpG addition to alum (AP or AH) resulted in a marginal trend of improved pNTs in stressed samples only, yet did not impact the storage stability profiles of IVX-411. In contrast, previous work with AH-formulations of a monomeric RBD antigen showed greatly improved immunogenicity and decreased stability upon CpG addition to alum. At elevated temperatures (25, 37°C), IVX-411 formulated with AH or AP displayed decreased in vitro stability compared to AV-formulated IVX-411and this rank-ordering correlated with in vivo performance (mouse pNT values). This case study highlights the importance of characterizing antigen-adjuvant interactions to develop low cost, aluminum-salt adjuvanted recombinant subunit vaccine candidates.
2023-10-01T00:00:00ZFormulation development and comparability studies with an aluminum-salt adjuvanted SARS-CoV-2 Spike ferritin nanoparticle vaccine antigen produced from two different cell lines (Dataset)Kumru, Ozan S.Sanyal, MrinmoyFriedland, NataliaHickey, JohnJoshi, RichaWeidenbacher, PaytonDo, JonathanCheng, Ya-ChenKim, Peter S.Joshi, Sangeeta B.Volkin, David B.https://hdl.handle.net/1808/347412023-08-21T15:38:28Z2023-01-01T00:00:00ZFormulation development and comparability studies with an aluminum-salt adjuvanted SARS-CoV-2 Spike ferritin nanoparticle vaccine antigen produced from two different cell lines (Dataset)
Kumru, Ozan S.; Sanyal, Mrinmoy; Friedland, Natalia; Hickey, John; Joshi, Richa; Weidenbacher, Payton; Do, Jonathan; Cheng, Ya-Chen; Kim, Peter S.; Joshi, Sangeeta B.; Volkin, David B.
The development of safe and effective second-generation COVID-19 vaccines to improve affordability and storage stability requirements remains a high priority to expand global coverage. In this report, we describe formulation development and comparability studies with a self-assembled SARS-CoV-2 spike ferritin nanoparticle vaccine antigen (called DCFHP), when produced in two different cell lines and formulated with an aluminum-salt adjuvant (Alhydrogel, AH). Varying levels of phosphate buffer altered the extent and strength of antigen-adjuvant interactions, and these formulations were evaluated for their (1) in vivo performance in mice and (2) in vitro stability profiles. Unadjuvanted DCFHP produced minimal immune responses while AH-adjuvanted formulations elicited greatly enhanced pseudovirus neutralization titers independent of ~100%, ~40% or ~10% of the DCFHP antigen adsorbed to AH. These formulations differed, however, in their in vitro stability properties as determined by biophysical studies and a competitive ELISA for measuring ACE2 receptor binding of AH-bound antigen. Interestingly, after one month of 4ºC storage, small increases in antigenicity with concomitant decreases in the ability to desorb the antigen from the AH were observed. Finally, we performed a comparability assessment of DCFHP antigen produced in Expi293 and CHO cells, which displayed expected differences in their N-linked oligosaccharide profiles. Despite consisting of different DCFHP glycoforms, these two preparations were highly similar in their key quality attributes including molecular size, structural integrity, conformational stability, binding to ACE2 receptor and mouse immunogenicity profiles. Taken together, these studies support future preclinical and clinical development of an AH-adjuvanted DCFHP vaccine candidate produced in CHO cells.
2023-01-01T00:00:00ZPhysicochemical characterization of biological and synthetic forms of two lipid A-based TLR4 agonistsHu, GangVarisco, David J.Das, SayanMiddaugh, C. RussellGardner, FrancescaErnst, Robert K.Picking, Wendy L.Picking, William D.https://hdl.handle.net/1808/347332023-08-15T06:07:20Z2023-07-08T00:00:00ZPhysicochemical characterization of biological and synthetic forms of two lipid A-based TLR4 agonists
Hu, Gang; Varisco, David J.; Das, Sayan; Middaugh, C. Russell; Gardner, Francesca; Ernst, Robert K.; Picking, Wendy L.; Picking, William D.
Toll-like receptor (TLR) agonists are recognized as potential immune-enhancing adjuvants and are included in several licensed vaccines. Monophosphoryl lipid A (MPL®, GlaxoSmithKline) is one such TLR4 agonist that has been approved for use in human vaccines, such as Cervarix and Shingrix. Due to the heterogeneous nature of biologically derived MPL and the need for safer and more potent adjuvants, our groups have developed the novel TLR4 agonist candidates, BECC438 and BECC470 using the Bacterial Enzymatic Combinatorial Chemistry (BECC) platform. BECC438 and BECC470 have been included in studies to test their adjuvant potential and found to be effective in vaccines against both viral and bacterial disease agents. Here, we report detailed biophysical characterization of BECC438 and BECC470 purified from a biological source (BECC438b and BECC470b, respectively) and synthesized chemically (BECC438s and BECC470s, respectively). Both BECC438s and BECC470s have identical acyl chain configurations, BECC438s is bis-phosphorylated and BECC470s is mono-phosphorylated with the removal of the 4′ phosphate moiety. We determined the phase transition temperatures for the acyl chains of BECC438b and BECC470b and found them to be different from those exhibited by their synthetic counterparts. Furthermore, the phosphate groups of BECC438b and BECC470b are more highly hydrated than are those of BECC438s and BECC470s. In addition to exploring the BECC molecules’ biophysical features in aqueous solution, we explored potential formulation of BECC438 and BECC470 with the aluminum-based adjuvant Alhydrogel and as part of an oil-in-water emulsion (Medimmune Emulsion or ME). All of the lipid A analogues could be fully absorbed to Alhydrogel or incorporated onto ME. Surprisingly, the BECC470s molecule, unlike the others, displayed a nearly baseline signal when monitored using a Limulus amebocyte lysate (LAL) endotoxin detection system. Despite this, it was shown to behave as an agonist for human and mouse TLR4 when tested using multiple cell-based systems. This work paves the way for further formulation optimization of two chemically defined TLR4 agonists that are showing great promise as vaccine adjuvants.
2023-07-08T00:00:00ZImmunogenicity and protective efficacy of nanoparticle formulations of L-SseB against Salmonella infectionDas, SayanHowlader, Debaki R.Lu, TiWhittier, Sean K.Hu, GangSharma, SimranDietz, Zackary K.Ratnakaram, Siva S. K.Varisco, David J.Ernst, Robert K.Picking, William D.Picking, Wendy L.https://hdl.handle.net/1808/347252023-08-15T06:07:37Z2023-06-30T00:00:00ZImmunogenicity and protective efficacy of nanoparticle formulations of L-SseB against Salmonella infection
Das, Sayan; Howlader, Debaki R.; Lu, Ti; Whittier, Sean K.; Hu, Gang; Sharma, Simran; Dietz, Zackary K.; Ratnakaram, Siva S. K.; Varisco, David J.; Ernst, Robert K.; Picking, William D.; Picking, Wendy L.
Salmonella enterica, a Gram-negative pathogen, has over 2500 serovars that infect a wide range of hosts. In humans, S. enterica causes typhoid or gastroenteritis and is a major public health concern. In this study, SseB (the tip protein of the Salmonella pathogenicity island 2 type III secretion system) was fused with the LTA1 subunit of labile-toxin from enterotoxigenic E. coli to make the self-adjuvanting antigen L-SseB. Two unique nanoparticle formulations were developed to allow multimeric presentation of L-SseB. Mice were vaccinated with these formulations and protective efficacy determined via challenging the mice with S. enterica serovars. The polysaccharide (chitosan) formulation was found to elicit better protection when compared to the squalene nanoemulsion. When the polysaccharide formulation was used to vaccinate rabbits, protection from S. enterica challenge was elicited. In summary, L-SseB in a particulate polysaccharide formulation appears to be an attractive candidate vaccine capable of broad protection against S. enterica.
2023-06-30T00:00:00Z