B21-High Throughput SAXS
I22-Small angle scattering & Diffraction
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Diamond Proposal Number(s):
[27906, 37100]
Abstract: The storage and distribution of vital protein therapeutics presents several complex challenges. Many medicines and vaccines need stable, temperature-controlled environments and chemical additives (excipients) such as preservatives to keep them effective and safe for use.
This requires cold storage infrastructure and reliable energy sources which not only puts the responsibility on the user but causes accessibility and affordability challenges, especially in developing countries where resources are limited.
Fig. 1
Fig. 1
Now researchers from the UK Universities of Manchester, Glasgow and Warwick have designed the world’s first hydrogel technology for the storage and distribution of crucial medicines and other biopharmaceuticals without the need for refrigeration or chemical additives. The aim is to provide more robust and equitable storage and delivery systems, benefitting everyone worldwide.
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Oct 2024
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John D.
Clarke
,
Helen M. E.
Duyvesteyn
,
Eva
Perez-Martin
,
Undīne
Latišenko
,
Claudine
Porta
,
Kathleen V.
Humphreys
,
Abigail L.
Hay
,
Jingshan
Ren
,
Elizabeth E.
Fry
,
Erwin
Van Den Born
,
Bryan
Charleston
,
Marie
Bonnet-Di Placido
,
Raymond J.
Owens
,
David I.
Stuart
,
John A.
Hammond
Open Access
Abstract: Foot-and-mouth disease vaccination using inactivated virus is suboptimal, as the icosahedral viral capsids often disassemble into antigenically distinct pentameric units during long-term storage, or exposure to elevated temperature or lowered pH, and thus raise a response that is no longer protective. Furthermore, as foot-and-mouth disease virus (FMDV)’s seven serotypes are antigenically diverse, cross-protection from a single serotype vaccine is limited, and most existing mouse and bovine antibodies and camelid single-domain heavy chain-only antibodies are serotype-specific. For quality control purposes, there is a real need for pan-serotype antibodies that clearly distinguish between pentamer (12S) and protective intact FMDV capsid. To date, few cross-serotype bovine-derived antibodies have been reported in the literature. We identify a bovine antibody with an ultralong CDR-H3, Ab117, whose structural analysis reveals that it binds to a deep, hydrophobic pocket on the interior surface of the capsid via the CDR-H3. Main-chain and hydrophobic interactions provide broad serotype specificity. ELISA analysis confirms that Ab117 is a novel pan-serotype and conformational epitope-specific 12S reagent, suitable for assessing capsid integrity.
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Oct 2024
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Krios I-Titan Krios I at Diamond
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Victoria A.
Avanzato
,
Trenton
Bushmaker
,
Kasopefoluwa Y.
Oguntuyo
,
Claude Kwe
Yinda
,
Helen M. E.
Duyvesteyn
,
Robert
Stass
,
Kimberly
Meade-White
,
Rebecca
Rosenke
,
Tina
Thomas
,
Neeltje
Van Doremalen
,
Greg
Saturday
,
Katie J.
Doores
,
Benhur
Lee
,
Thomas A.
Bowden
,
Vincent J.
Munster
Diamond Proposal Number(s):
[20223]
Abstract: Nipah virus (NiV) is a highly pathogenic paramyxovirus capable of causing severe respiratory and neurologic disease in humans. Currently, there are no licensed vaccines or therapeutics against NiV, underscoring the urgent need for the development of countermeasures. The NiV surface-displayed glycoproteins, NiV-G and NiV-F, mediate host cell attachment and fusion, respectively, and are heavily targeted by host antibodies. Here, we describe a vaccination-derived neutralizing monoclonal antibody, mAb92, that targets NiV-F. Structural characterization of the Fab region bound to NiV-F (NiV-F–Fab92) by cryo-electron microscopy analysis reveals an epitope in the DIII domain at the membrane distal apex of NiV-F, an established site of vulnerability on the NiV surface. Further, prophylactic treatment of hamsters with mAb92 offered complete protection from NiV disease, demonstrating beneficial activity of mAb92 in vivo. This work provides support for targeting NiV-F in the development of vaccines and therapeutics against NiV.
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Sep 2024
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I04-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography
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Diamond Proposal Number(s):
[23459]
Open Access
Abstract: There is an urgent need for improved malaria vaccine immunogens. Invasion of erythrocytes by Plasmodium falciparum is essential for its life cycle, preceding symptoms of disease and parasite transmission. Antibodies which target PfRH5 are highly effective at preventing erythrocyte invasion and the most potent growth-inhibitory antibodies bind a single epitope. Here we use structure-guided approaches to design a small synthetic immunogen, RH5-34EM which recapitulates this epitope. Structural biology and biophysics demonstrate that RH5-34EM is correctly folded and binds neutralising monoclonal antibodies with nanomolar affinity. In immunised rats, RH5-34EM induces PfRH5-targeting antibodies that inhibit parasite growth. While PfRH5-specific antibodies were induced at a lower concentration by RH5-34EM than by PfRH5, RH5-34EM induced antibodies that were a thousand-fold more growth-inhibitory as a factor of PfRH5-specific antibody concentration. Finally, we show that priming with RH5-34EM and boosting with PfRH5 achieves the best balance between antibody quality and quantity and induces the most effective growth-inhibitory response. This rationally designed vaccine immunogen is now available for use as part of future malaria vaccines, alone or in combination with other immunogens.
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Sep 2024
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Krios IV-Titan Krios IV at Diamond
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Diamond Proposal Number(s):
[24039]
Open Access
Abstract: Despite treatment and other interventions, an effective prophylactic HIV vaccine is still an essential goal in the control of HIV. Inducing robust and long-lasting antibody responses is one of the main targets of an HIV vaccine. The delivery of HIV envelope glycoproteins (Env) using nanoparticle (NP) platforms has been shown to elicit better immunogenicity than soluble HIV Env. In this paper, we describe the development of a nanoparticle-based vaccine decorated with HIV Env using the SpyCatcher/SpyTag system. The Env utilised in this study, CAP255, was derived from a transmitted founder virus isolated from a patient who developed broadly neutralising antibodies. Negative stain and cryo-electron microscopy analyses confirmed the assembly and stability of the mi3 into uniform icosahedral NPs surrounded by regularly spaced CAP255 gp140 Env trimers. A three-dimensional reconstruction of CAP255 gp140 SpyTag–SpyCatcher mi3 clearly showed Env trimers projecting from the centre of each of the pentagonal dodecahedral faces of the NP. To our knowledge, this is the first study to report the formation of SpyCatcher pentamers on the dodecahedral faces of mi3 NPs. To investigate the immunogenicity, rabbits were primed with two doses of DNA vaccines expressing the CAP255 gp150 and a mosaic subtype C Gag and boosted with three doses of the NP-developed autologous Tier 2 CAP255 neutralising antibodies (Nabs) and low levels of heterologous CAP256SU NAbs.
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Sep 2024
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I04-Macromolecular Crystallography
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Michelle R.
Garnsey
,
Matthew C.
Robinson
,
Luong T.
Nguyen
,
Rhonda
Cardin
,
Joseph
Tillotson
,
Ellene
Mashalidis
,
Aijia
Yu
,
Lisa
Aschenbrenner
,
Amanda
Balesano
,
Amin
Behzadi
,
Britton
Boras
,
Jeanne S.
Chang
,
Heather
Eng
,
Andrew
Ephron
,
Tim
Foley
,
Kristen K.
Ford
,
James M.
Frick
,
Scott
Gibson
,
Li
Hao
,
Brett
Hurst
,
Amit S.
Kalgutkar
,
Magdalena
Korczynska
,
Zsofia
Lengyel-Zhand
,
Liping
Gao
,
Hannah R.
Meredith
,
Nandini C.
Patel
,
Jana
Polivkova
,
Devendra
Rai
,
Colin R.
Rose
,
Hussin
Rothan
,
Sylvie K.
Sakata
,
Thomas R.
Vargo
,
Wenying
Qi
,
Huixian
Wu
,
Yiping
Liu
,
Irina
Yurgelonis
,
Jinzhi
Zhang
,
Yuao
Zhu
,
Lei
Zhang
,
Alpha A.
Lee
Open Access
Abstract: Vaccines and first-generation antiviral therapeutics have provided important protection against COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, there remains a need for additional therapeutic options that provide enhanced efficacy and protection against potential viral resistance. The SARS-CoV-2 papain-like protease (PLpro) is one of the two essential cysteine proteases involved in viral replication. While inhibitors of the SARS-CoV-2 main protease have demonstrated clinical efficacy, known PLpro inhibitors have, to date, lacked the inhibitory potency and requisite pharmacokinetics to demonstrate that targeting PLpro translates to in vivo efficacy in a preclinical setting. Here, we report the machine learning–driven discovery of potent, selective, and orally available SARS-CoV-2 PLpro inhibitors, with lead compound PF-07957472 (4) providing robust efficacy in a mouse-adapted model of COVID-19 infection.
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Aug 2024
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B21-High Throughput SAXS
I22-Small angle scattering & Diffraction
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Diamond Proposal Number(s):
[27906, 37100]
Open Access
Abstract: A long-standing challenge is how to formulate proteins and vaccines to retain function during storage and transport and to remove the burdens of cold-chain management. Any solution must be practical to use, with the protein being released or applied using clinically relevant triggers. Advanced biologic therapies are distributed cold, using substantial energy, limiting equitable distribution in low-resource countries and placing responsibility on the user for correct storage and handling. Cold-chain management is the best solution at present for protein transport but requires substantial infrastructure and energy. For example, in research laboratories, a single freezer at −80 °C consumes as much energy per day as a small household1. Of biological (protein or cell) therapies and all vaccines, 75% require cold-chain management; the cost of cold-chain management in clinical trials has increased by about 20% since 2015, reflecting this complexity. Bespoke formulations and excipients are now required, with trehalose2, sucrose or polymers3 widely used, which stabilize proteins by replacing surface water molecules and thereby make denaturation thermodynamically less likely; this has enabled both freeze-dried proteins and frozen proteins. For example, the human papilloma virus vaccine requires aluminium salt adjuvants to function, but these render it unstable against freeze–thaw4, leading to a very complex and expensive supply chain. Other ideas involve ensilication5 and chemical modification of proteins6. In short, protein stabilization is a challenge with no universal solution7,8. Here we designed a stiff hydrogel that stabilizes proteins against thermal denaturation even at 50 °C, and that can, unlike present technologies, deliver pure, excipient-free protein by mechanically releasing it from a syringe. Macromolecules can be loaded at up to 10 wt% without affecting the mechanism of release. This unique stabilization and excipient-free release synergy offers a practical, scalable and versatile solution to enable the low-cost, cold-chain-free and equitable delivery of therapies worldwide.
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Jul 2024
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Open Access
Abstract: Spike proteins protrude from the SARS-CoV-2 viral envelope and are responsible for initiating fusion into human epithelial cells by binding Angiotensin-Converting Enzyme 2 receptors on the host cell surface. Due to their exposed location on the outside of the virion and their key role in infection, SARS-CoV-2 spike proteins are an important target for vaccine development and drug design. Over the last two years, many spike protein structures have been experimentally determined, providing essential details into the complex structural rearrangements that occur after receptor binding and during fusion of the virion with the host cell, as well as into the interactions of spike protein molecules with antibodies. SARS-CoV-2 variants, particularly those associated with reduced vaccine efficacy, are strongly associated with mutations in two domains of the SARS-CoV-2 spike protein, namely the receptor binding domain and the N-terminal domain, which have both been structurally characterized. This review provides a comprehensive overview of the structural knowledge acquired over the past four years on the SARS-CoV-2 spike protein and its critical role in viral infection.
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Jul 2024
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I04-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography
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Lawrence T.
Wang
,
Andrew J. R.
Cooper
,
Brendan
Farrell
,
Kazutoyo
Miura
,
Ababacar
Diouf
,
Nicole
Müller-Sienerth
,
Cécile
Crosnier
,
Lauren
Purser
,
Payton J.
Kirtley
,
Maciej
Maciuszek
,
Jordan R.
Barrett
,
Kirsty
Mchugh
,
Rodney
Ogwang
,
Courtney
Tucker
,
Shanping
Li
,
Safiatou
Doumbo
,
Didier
Doumtabe
,
Chul-Woo
Pyo
,
Jeff
Skinner
,
Carolyn M.
Nielsen
,
Sarah E.
Silk
,
Kassoum
Kayentao
,
Aissata
Ongoiba
,
Ming
Zhao
,
Doan C.
Nguyen
,
F. Eun-Hyung
Lee
,
Angela M.
Minassian
,
Daniel E.
Geraghty
,
Boubacar
Traore
,
Robert A.
Seder
,
Brandon K.
Wilder
,
Peter D.
Crompton
,
Gavin J.
Wright
,
Carole A.
Long
,
Simon J.
Draper
,
Matthew K.
Higgins
,
Joshua
Tan
Open Access
Abstract: Plasmodium falciparum reticulocyte-binding protein homolog 5 (RH5) is the most advanced blood-stage malaria vaccine candidate and is being evaluated for efficacy in endemic regions, emphasizing the need to study the underlying antibody response to RH5 during natural infection, which could augment or counteract responses to vaccination. Here, we found that RH5-reactive B cells were rare, and circulating immunoglobulin G (IgG) responses to RH5 were short-lived in malaria-exposed Malian individuals, despite repeated infections over multiple years. RH5-specific monoclonal antibodies isolated from eight malaria-exposed individuals mostly targeted non-neutralizing epitopes, in contrast to antibodies isolated from five RH5-vaccinated, malaria-naive UK individuals. However, MAD8–151 and MAD8–502, isolated from two malaria-exposed Malian individuals, were among the most potent neutralizers out of 186 antibodies from both cohorts and targeted the same epitopes as the most potent vaccine-induced antibodies. These results suggest that natural malaria infection may boost RH5-vaccine-induced responses and provide a clear strategy for the development of next-generation RH5 vaccines.
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Jul 2024
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I03-Macromolecular Crystallography
I04-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography
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Jordan R.
Barrett
,
Dimitra
Pipini
,
Nathan D.
Wright
,
Andrew J. R.
Cooper
,
Giacomo
Gorini
,
Doris
Quinkert
,
Amelia M.
Lias
,
Hannah
Davies
,
Cassandra A.
Rigby
,
Maya
Aleshnick
,
Barnabas G.
Williams
,
William J.
Bradshaw
,
Neil G.
Paterson
,
Thomas
Martinson
,
Payton
Kirtley
,
Luc
Picard
,
Christine D.
Wiggins
,
Francesca R.
Donnellan
,
Lloyd D. W.
King
,
Lawrence T.
Wang
,
Jonathan F.
Popplewell
,
Sarah E.
Silk
,
Jed
De Ruiter Swain
,
Katherine
Skinner
,
Vinayaka
Kotraiah
,
Amy R.
Noe
,
Randall S.
Macgill
,
C. Richter
King
,
Ashley J.
Birkett
,
Lorraine A.
Soisson
,
Angela M.
Minassian
,
Douglas A.
Lauffenburger
,
Kazutoyo
Miura
,
Carole A.
Long
,
Brandon K.
Wilder
,
Lizbe
Koekemoer
,
Joshua
Tan
,
Carolyn M.
Nielsen
,
Kirsty
Mchugh
,
Simon J.
Draper
Diamond Proposal Number(s):
[28172]
Open Access
Abstract: The highly conserved and essential Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has emerged as the leading target for vaccines against the disease-causing blood stage of malaria. However, the features of the human vaccine-induced antibody response that confer highly potent inhibition of malaria parasite invasion into red blood cells are not well defined. Here, we characterize 236 human IgG monoclonal antibodies, derived from 15 donors, induced by the most advanced PfRH5 vaccine. We define the antigenic landscape of this molecule and establish that epitope specificity, antibody association rate, and intra-PfRH5 antibody interactions are key determinants of functional anti-parasitic potency. In addition, we identify a germline IgG gene combination that results in an exceptionally potent class of antibody and demonstrate its prophylactic potential to protect against P. falciparum parasite challenge in vivo. This comprehensive dataset provides a framework to guide rational design of next-generation vaccines and prophylactic antibodies to protect against blood-stage malaria.
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Jul 2024
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