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Article ; Online: Engineered antibody fusion proteins for targeted disease therapy.

Silver, Aliyah B / Leonard, Elissa K / Gould, Joseph R / Spangler, Jamie B

2021 Volume 42, Issue 12, Page(s) 1064–1081

Abstract: Since the FDA approval of the first therapeutic antibody 35 years ago, antibody-based products have gained prominence in the pharmaceutical market. Building on the early successes of monoclonal antibodies, more recent efforts have capitalized on the ... ...

Abstract	Since the FDA approval of the first therapeutic antibody 35 years ago, antibody-based products have gained prominence in the pharmaceutical market. Building on the early successes of monoclonal antibodies, more recent efforts have capitalized on the exquisite specificity and/or favorable pharmacokinetic properties of antibodies by developing fusion proteins that enable targeted delivery of therapeutic payloads which are otherwise ineffective when administered systemically. This review focuses on recent engineering and translational advances for therapeutics that genetically fuse antibodies to disease-relevant payloads, including cytokines, toxins, enzymes, neuroprotective agents, and soluble factor traps. With numerous antibody fusion proteins in the clinic and other innovative molecules poised to follow suit, these potent, multifunctional drug candidates promise to be a major player in the therapeutic development landscape for years to come.
MeSH term(s)	Antibodies, Monoclonal ; Cytokines ; Humans
Chemical Substances	Antibodies, Monoclonal ; Cytokines
Language	English
Publishing date	2021-10-25
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	282846-7
ISSN	1873-3735 ; 0165-6147
ISSN (online)	1873-3735
ISSN	0165-6147
DOI	10.1016/j.tips.2021.09.009
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Article: Antibody–Invertase Fusion Protein Enables Quantitative Detection of SARS-CoV-2 Antibodies Using Widely Available Glucometers

Leonard, Elissa K. / Aller Pellitero, Miguel / Juelg, Boris / Spangler, Jamie B. / Arroyo-Currás, Netzahualcóyotl

Journal of the American Chemical Society. 2022 June 08, v. 144, no. 25

2022

Abstract: Rapid diagnostics that can accurately inform patients of disease risk and protection are critical to mitigating the spread of the current COVID-19 pandemic and future infectious disease outbreaks. To be effective, such diagnostics must rely on simple, ... ...

Abstract	Rapid diagnostics that can accurately inform patients of disease risk and protection are critical to mitigating the spread of the current COVID-19 pandemic and future infectious disease outbreaks. To be effective, such diagnostics must rely on simple, cost-effective, and widely available equipment and should be compatible with existing telehealth infrastructure to facilitate data access and remote care. Commercial glucometers are an established detection technology that can overcome the cost, time, and trained personnel requirements of current benchtop-based antibody serology assays when paired with reporter molecules that catalyze glucose conversion. To this end, we developed an enzymatic reporter that, when bound to disease-specific patient antibodies, produces glucose in proportion to the level of antibodies present in the patient sample. Although a straightforward concept, the coupling of enzymatic reporters to secondary antibodies or antigens often results in low yields, indeterminant stoichiometry, reduced target binding, and poor catalytic efficiency. Our enzymatic reporter is a novel fusion protein that comprises an antihuman immunoglobulin G (IgG) antibody genetically fused to two invertase molecules. The resulting fusion protein retains the binding affinity and catalytic activity of the constituent proteins and serves as an accurate reporter for immunoassays. Using this fusion, we demonstrate quantitative glucometer-based measurement of anti-SARS-CoV-2 spike protein antibodies in blinded clinical sample training sets. Our results demonstrate the ability to detect SARS-CoV-2-specific IgGs in patient serum with precise agreement to benchmark commercial immunoassays. Because our fusion protein binds all human IgG isotypes, it represents a versatile tool for detection of disease-specific antibodies in a broad range of biomedical applications.
Keywords	COVID-19 infection ; Severe acute respiratory syndrome coronavirus 2 ; antibodies ; beta-fructofuranosidase ; blood serum ; catalytic activity ; cost effectiveness ; diagnostic techniques ; equipment ; glucose ; human resources ; humans ; immunoglobulin G ; infrastructure ; patients ; risk ; serology ; stoichiometry ; telemedicine
Language	English
Dates of publication	2022-0608
Size	p. 11226-11237.
Publishing place	American Chemical Society
Document type	Article
ZDB-ID	3155-0
ISSN	1520-5126 ; 0002-7863
ISSN (online)	1520-5126
ISSN	0002-7863
DOI	10.1021/jacs.2c02537
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Article ; Online: Antibody-Invertase Fusion Protein Enables Quantitative Detection of SARS-CoV-2 Antibodies Using Widely Available Glucometers.

Leonard, Elissa K / Aller Pellitero, Miguel / Juelg, Boris / Spangler, Jamie B / Arroyo-Currás, Netzahualcóyotl

Journal of the American Chemical Society

2022 Volume 144, Issue 25, Page(s) 11226–11237

Abstract	Rapid diagnostics that can accurately inform patients of disease risk and protection are critical to mitigating the spread of the current COVID-19 pandemic and future infectious disease outbreaks. To be effective, such diagnostics must rely on simple, cost-effective, and widely available equipment and should be compatible with existing telehealth infrastructure to facilitate data access and remote care. Commercial glucometers are an established detection technology that can overcome the cost, time, and trained personnel requirements of current benchtop-based antibody serology assays when paired with reporter molecules that catalyze glucose conversion. To this end, we developed an enzymatic reporter that, when bound to disease-specific patient antibodies, produces glucose in proportion to the level of antibodies present in the patient sample. Although a straightforward concept, the coupling of enzymatic reporters to secondary antibodies or antigens often results in low yields, indeterminant stoichiometry, reduced target binding, and poor catalytic efficiency. Our enzymatic reporter is a novel fusion protein that comprises an antihuman immunoglobulin G (IgG) antibody genetically fused to two invertase molecules. The resulting fusion protein retains the binding affinity and catalytic activity of the constituent proteins and serves as an accurate reporter for immunoassays. Using this fusion, we demonstrate quantitative glucometer-based measurement of anti-SARS-CoV-2 spike protein antibodies in blinded clinical sample training sets. Our results demonstrate the ability to detect SARS-CoV-2-specific IgGs in patient serum with precise agreement to benchmark commercial immunoassays. Because our fusion protein binds all human IgG isotypes, it represents a versatile tool for detection of disease-specific antibodies in a broad range of biomedical applications.
MeSH term(s)	Antibodies, Viral ; COVID-19/diagnosis ; Glucose ; Humans ; Immunoglobulin G ; Pandemics ; SARS-CoV-2 ; Sensitivity and Specificity ; beta-Fructofuranosidase
Chemical Substances	Antibodies, Viral ; Immunoglobulin G ; beta-Fructofuranosidase (EC 3.2.1.26) ; Glucose (IY9XDZ35W2)
Language	English
Publishing date	2022-06-08
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	3155-0
ISSN	1520-5126 ; 0002-7863
ISSN (online)	1520-5126
ISSN	0002-7863
DOI	10.1021/jacs.2c02537
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Article ; Online: Weaponizing T-cell receptors through molecular engineering.

Leonard, Elissa K / Leff, Michael I / Spangler, Jamie B

The Journal of biological chemistry

2019 Volume 294, Issue 15, Page(s) 5805–5806

Abstract: T-cell receptors (TCRs) recognize pathogens to ignite immune responses, making them attractive scaffolds for development as immunotherapeutics. However, manipulation of TCRs has been impeded by difficulties in their engineering and expression. Wagner and ...

Abstract	T-cell receptors (TCRs) recognize pathogens to ignite immune responses, making them attractive scaffolds for development as immunotherapeutics. However, manipulation of TCRs has been impeded by difficulties in their engineering and expression. Wagner and colleagues now establish new platforms to generate high-affinity TCR variants that potently activate T cells, and they also create soluble TCR fusion proteins that specifically recognize cognate peptides. This work provides specific tools to combat cytomegalovirus (CMV) infection and helps illuminate a general path to actuation of engineered TCR-based therapeutics.
MeSH term(s)	Animals ; Gene Expression ; Humans ; Protein Engineering ; Receptors, Antigen, T-Cell/genetics ; Solubility
Chemical Substances	Receptors, Antigen, T-Cell
Language	English
Publishing date	2019-04-12
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.H119.008479
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Article: Engineered cytokine/antibody fusion proteins improve delivery of IL-2 to pro-inflammatory cells and promote antitumor activity.

Leonard, Elissa K / Tomala, Jakub / Gould, Joseph R / Leff, Michael I / Lin, Jian-Xin / Li, Peng / Porter, Mitchell J / Johansen, Eric R / Thompson, Ladaisha / Cao, Shanelle D / Henclova, Tereza / Huliciak, Maros / Vaněk, Ondřej / Kovar, Marek / Leonard, Warren J / Spangler, Jamie B

bioRxiv : the preprint server for biology

2023

Abstract: Progress in cytokine engineering is driving therapeutic translation by overcoming the inherent limitations of these proteins as drugs. The interleukin-2 (IL-2) cytokine harbors great promise as an immune stimulant for cancer treatment. However, the ... ...

Abstract	Progress in cytokine engineering is driving therapeutic translation by overcoming the inherent limitations of these proteins as drugs. The interleukin-2 (IL-2) cytokine harbors great promise as an immune stimulant for cancer treatment. However, the cytokine's concurrent activation of both pro-inflammatory immune effector cells and anti-inflammatory regulatory T cells, its toxicity at high doses, and its short serum half-life have limited clinical application. One promising approach to improve the selectivity, safety, and longevity of IL-2 is complexation with anti-IL-2 antibodies that bias the cytokine towards the activation of immune effector cells (i.e., effector T cells and natural killer cells). Although this strategy shows therapeutic potential in preclinical cancer models, clinical translation of a cytokine/antibody complex is complicated by challenges in formulating a multi-protein drug and concerns about complex stability. Here, we introduce a versatile approach to designing intramolecularly assembled single-agent fusion proteins (immunocytokines, ICs) comprising IL-2 and a biasing anti-IL-2 antibody that directs the cytokine's activities towards immune effector cells. We establish the optimal IC construction and further engineer the cytokine/antibody affinity to improve immune biasing function. We demonstrate that our IC preferentially activates and expands immune effector cells, leading to superior antitumor activity compared to natural IL-2 without inducing toxicities associated with IL-2 administration. Collectively, this work presents a roadmap for the design and translation of immunomodulatory cytokine/antibody fusion proteins.
Language	English
Publishing date	2023-05-04
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.05.03.539272
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Article ; Online: In Vivo Stimulation of Therapeutic Antigen-Specific T Cells in an Artificial Lymph Node Matrix.

Advanced materials (Deerfield Beach, Fla.)

2024 , Page(s) e2310043

Abstract: T cells are critical mediators of antigen-specific immune responses and are common targets for immunotherapy. Biomaterial scaffolds have previously been used to stimulate antigen-presenting cells to elicit antigen-specific immune responses; however, ... ...

Abstract	T cells are critical mediators of antigen-specific immune responses and are common targets for immunotherapy. Biomaterial scaffolds have previously been used to stimulate antigen-presenting cells to elicit antigen-specific immune responses; however, structural and molecular features that directly stimulate and expand naïve, endogenous, tumor-specific T cells in vivo have not been defined. Here, an artificial lymph node (aLN) matrix is created, which consists of an extracellular matrix hydrogel conjugated with peptide-loaded-MHC complex (Signal 1), the co-stimulatory signal anti-CD28 (Signal 2), and a tethered IL-2 (Signal 3), that can bypass challenges faced by other approaches to activate T cells in situ such as vaccines. This dynamic immune-stimulating platform enables direct, in vivo antigen-specific CD8+ T cell stimulation, as well as recruitment and coordination of host immune cells, providing an immuno-stimulatory microenvironment for antigen-specific T cell activation and expansion. Co-injecting the aLN with naïve, wild-type CD8+ T cells results in robust activation and expansion of tumor-targeted T cells that kill target cells and slow tumor growth in several distal tumor models. The aLN platform induces potent in vivo antigen-specific CD8+ T cell stimulation without the need for ex vivo priming or expansion and enables in situ manipulation of antigen-specific responses for immunotherapies.
Language	English
Publishing date	2024-02-15
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1474949-X
ISSN	1521-4095 ; 0935-9648
ISSN (online)	1521-4095
ISSN	0935-9648
DOI	10.1002/adma.202310043
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Article ; Online: Nanoparticle-based modulation of CD4

Isser, Ariel / Silver, Aliyah B / Pruitt, Hawley C / Mass, Michal / Elias, Emma H / Aihara, Gohta / Kang, Si-Sim / Bachmann, Niklas / Chen, Ying-Yu / Leonard, Elissa K / Bieler, Joan G / Chaisawangwong, Worarat / Choy, Joseph / Shannon, Sydney R / Gerecht, Sharon / Weber, Jeffrey S / Spangler, Jamie B / Schneck, Jonathan P

Nature communications

2022 Volume 13, Issue 1, Page(s) 6086

Abstract: ... Helper ( ... ...

Abstract	Helper (CD4
MeSH term(s)	Animals ; Antigen-Presenting Cells ; CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; HLA Antigens ; Humans ; Immunotherapy, Adoptive ; Mice ; Nanoparticles
Chemical Substances	HLA Antigens
Language	English
Publishing date	2022-10-14
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-022-33597-y
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Article ; Online: Supramolecular filaments for concurrent ACE2 docking and enzymatic activity silencing enable coronavirus capture and infection prevention.

Anderson, Caleb F / Wang, Qiong / Stern, David / Leonard, Elissa K / Sun, Boran / Fergie, Kyle J / Choi, Chang-Yong / Spangler, Jamie B / Villano, Jason / Pekosz, Andrew / Brayton, Cory F / Jia, Hongpeng / Cui, Honggang

Matter

2022 Volume 6, Issue 2, Page(s) 583–604

Abstract: Coronaviruses have historically precipitated global pandemics of severe acute respiratory syndrome (SARS) into devastating public health crises. Despite the virus's rapid rate of mutation, all SARS coronavirus 2 (SARS-CoV-2) variants are known to gain ... ...

Abstract	Coronaviruses have historically precipitated global pandemics of severe acute respiratory syndrome (SARS) into devastating public health crises. Despite the virus's rapid rate of mutation, all SARS coronavirus 2 (SARS-CoV-2) variants are known to gain entry into host cells primarily through complexation with angiotensin-converting enzyme 2 (ACE2). Although ACE2 has potential as a druggable decoy to block viral entry, its clinical use is complicated by its essential biological role as a carboxypeptidase and hindered by its structural and chemical instability. Here we designed supramolecular filaments, called fACE2, that can silence ACE2's enzymatic activity and immobilize ACE2 to their surface through enzyme-substrate complexation. This docking strategy enables ACE2 to be effectively delivered in inhalable aerosols and improves its structural stability and functional preservation. fACE2 exhibits enhanced and prolonged inhibition of viral entry compared with ACE2 alone while mitigating lung injury
Language	English
Publishing date	2022-12-12
Publishing country	United States
Document type	Journal Article
ISSN	2590-2385
ISSN (online)	2590-2385
DOI	10.1016/j.matt.2022.11.027
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Article ; Online: Recurrence of severe diverticulitis is associated with age and birth decade.

Leonard, Molly E / Horns, Joshua J / Allen-Brady, Kristina / Ozanne, Elissa M / Wallace, Andrea S / Brooke, Benjamin S / Supiano, Mark A / Cohan, Jessica N

Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract

2024 Volume 28, Issue 4, Page(s) 507–512

Abstract: Background: The risk of recurrence is an important consideration when deciding to treat patients medically or with elective colectomy after recovery from diverticulitis. It is unclear whether age is associated with recurrence. This study aimed to ... ...

Abstract	Background: The risk of recurrence is an important consideration when deciding to treat patients medically or with elective colectomy after recovery from diverticulitis. It is unclear whether age is associated with recurrence. This study aimed to examine the relationship between age and the risk of recurrent diverticulitis while considering important epidemiologic factors, such as birth decade. Methods: The Utah Population Database was used to identify individuals with incident severe diverticulitis, defined as requiring an emergency department visit or hospitalization, between 1998 and 2018. This study measured the relationship between age and recurrent severe diverticulitis after adjusting for birth decade and other important variables, such as sex, urban/rural status, complicated diverticulitis, and body mass index using a Cox proportional hazards model. Results: The cohort included 8606 individuals with a median age of 61 years at index diverticulitis diagnosis. After adjustment, among individuals born in the same birth decade, increasing age at diverticulitis onset was associated with an increased risk of recurrent diverticulitis (hazard ratio [HR] for 10 years, 1.8; 95% CI, 1.5-2.1). Among individuals with the same age of onset, those born in a more recent birth decade were also at greater risk of recurrent diverticulitis (HR, 1.9; 95% CI, 1.6-2.3). Conclusion: Among individuals with an index episode of severe diverticulitis, recurrence was associated with increasing age and more recent birth decade. Clinicians may wish to employ age-specific strategies when counseling patients regarding treatment options after a diverticulitis diagnosis.
MeSH term(s)	Humans ; Middle Aged ; Child ; Diverticulitis, Colonic/epidemiology ; Diverticulitis, Colonic/surgery ; Diverticulitis, Colonic/complications ; Retrospective Studies ; Diverticulitis/complications ; Hospitalization ; Colectomy/adverse effects ; Recurrence
Language	English
Publishing date	2024-02-09
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2012365-6
ISSN	1873-4626 ; 1934-3213 ; 1091-255X
ISSN (online)	1873-4626 ; 1934-3213
ISSN	1091-255X
DOI	10.1016/j.gassur.2023.12.028
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Article ; Online: Design and validation of a corneal bioreactor.

Leonard, Elissa K / Pai, Vincent H / Amberg, Philip / Gardner, Jens / Orwin, Elizabeth J

Biotechnology and bioengineering

2012 Volume 109, Issue 12, Page(s) 3189–3198

Abstract: Mechanical strain is an important signal that influences the behavior and properties of cells in a wide variety of tissues. Physiologically similar mechanical strain can revert cultured cells to a more normal phenotype. Here, we have demonstrated that 3% ...

Abstract	Mechanical strain is an important signal that influences the behavior and properties of cells in a wide variety of tissues. Physiologically similar mechanical strain can revert cultured cells to a more normal phenotype. Here, we have demonstrated that 3% equibiaxial (EB) and uniaxial strains confer favorable protein expression in cultured rabbit corneal fibroblasts (RCFs), with approximately 35% and 65% reduction in expression of α-smooth muscle actin (α-SMA), respectively. We have designed a novel bioreactor that is capable of imparting up to 7% EB strain and up to 6% EB strain using a cornea-shaped post. Additional features of the bioreactor include the application of shear stress to cells in culture and the ability to image cells using optical coherence microscopy (OCM) without being removed from the system.
MeSH term(s)	Actins ; Animals ; Bioreactors ; Cell Culture Techniques/methods ; Cornea/cytology ; Fibroblasts/cytology ; Fibroblasts/metabolism ; Fibroblasts/physiology ; Finite Element Analysis ; Rabbits ; Reproducibility of Results ; Stress, Mechanical ; Tissue Engineering/methods ; Tomography, Optical Coherence
Chemical Substances	Actins
Language	English
Publishing date	2012-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	280318-5
ISSN	1097-0290 ; 0006-3592
ISSN (online)	1097-0290
ISSN	0006-3592
DOI	10.1002/bit.24587
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