As atrocity has become characteristic of modern history, testimonial writing has become a major twentieth-century genre. Untimely Interventions relates testimonial writing, or witnessing, to the ...cultural situation of aftermath, exploring ways in which a culture can be haunted by its own history.
Ross Chambers argues that culture produces itself as civilized by denying the forms of collective violence and other traumatic experience that it cannot control. In the context of such denial, personal accounts of collective disaster can function as a form of counter-denial. By investigating a range of writing on AIDS, the First World War, and the Holocaust, Chambers shows how such writing produces a rhetorical effect of haunting, as it seeks to describe the reality of those experiences culture renders unspeakable.
Ross Chambers is Professor of Romance Languages at the University of Michigan. His other books includeFacing It: AIDS Diaries and the Death of the Author.
To exploit highly conserved and difficult drug targets, including multipass membrane proteins, monoclonal antibody discovery efforts increasingly rely on the advantages offered by divergent species ...such as rabbits, camelids, and chickens. Here, we provide an overview of antibody discovery technologies, analyze gaps in therapeutic antibodies that stem from the historic use of mice, and examine opportunities to exploit previously inaccessible targets through discovery now possible in alternate species. We summarize the clinical development of antibodies raised from divergent species, discussing how these animals enable robust immune responses against highly conserved binding sites and yield antibodies capable of penetrating functional pockets via long HCDR3 regions. We also discuss the value of pan-reactive molecules often produced by these hosts, and how these antibodies can be tested in accessible animal models, offering a faster path to clinical development.
What happens to poetic beauty when history turns the poet from one who contemplates natural beauty and the sublime to one who attempts to reconcile the practice of art with the hustle and noise of ...the city? An Atmospherics of the City traces Charles Baudelaire's evolution from a writer who practices a form of fetishizing aesthetics in which poetry works to beautify the ordinary to one who perceives background noise and disorder-the city's version of a transcendent atmosphere-as evidence of the malign work of a transcendent god of time, history, and ultimate destruction. Analyzing this shift, particularly as evidenced in Tableaux parisiens and Le Spleen de Paris, Ross Chambers shows how Baudelaire's disenchantment with the politics of his day and the coincident rise of overpopulation, poverty, and Haussmann's modernization of Paris influenced the poet's work to conceive a poetry of allegory, one with the power to alert and disalienate its otherwise inattentive reader whose senses have long been dulled by the din of his environment. Providing a completely new and original understanding of both Baudelaire's ethics and his aesthetics, Chambers reveals how the shift from themes of the supernatural in Baudelaire to ones of alienation allowed a new way for him to articulate and for his fellow Parisians to comprehend the rapidly changing conditions of the city and, in the process, to invent a "modern beauty" from the realm of suffering and the abject as they embodied forms of urban experience.
There's an interesting essay by Roger Chartier, in which he shows that European universities, in the seventeenth and eighteenth centuries, began to take in more students than the Church, with its ...monopoly on classical knowledge, could absorb.1 There began to be a surplus of educated men for whom there were insufficient positions as priests, curates and schoolmasters; and thus the modern category of the intellectual began to emerge, which in turn made a major contribution to the Enlightenment movement, and with it to the social disturbances that finally brought about the end of the ancien regime and the emergence out of revolution of the modern world. Out of such small disorders in the best - regulated systems - that is to say, out of the vulnerability of social conventions to the entropy that inevitably and parasitically inhabits them - large social transformations can emerge, only to be subject in due course to more disorder, and to further changes.
BackgroundGPRC5D is a G protein-coupled receptor that is expressed on multiple myeloma cells but absent from most healthy tissues except for hair follicles. Tumor-associated antigens like GPRC5D with ...little or no expression in healthy tissue are attractive targets for potent anti-tumor modalities including T cell-engaging bispecific antibodies. There is immense interest in GPRC5D as a multiple myeloma target, but currently no clinically approved therapies against it.MethodsMultipass membrane proteins are valuable therapeutic targets in oncology and other disease areas but are largely inaccessible as antibody targets due to their poor expression, membrane-dependent structure, small extracellular regions, and poor immunogenicity due to sequence conservation. We developed an antibody discovery platform (MPS) that specifically addresses each of these challenges. This platform utilizes advanced immunization techniques including DNA, mRNA, and Lipoparticles (virus-like particles). It also employs chickens as an evolutionarily divergent host species for robust immune responses against conserved targets. Antibodies raised in chickens are directly humanized prior to isolation reducing the need for downstream engineering. From the parental antibodies isolated, we engineered panels of GPRC5DxCD3 bispecific antibodies using multiple formats and CD3 arms, that encompass different geometries and binding stoichiometries, as these factors are expected to play a critical role for in vitro and in vivo potency.ResultsWe immunized chickens with GPRC5D and obtained high-titer immune responses. A subset of antibodies from this discovery program was configured as bispecific molecules using a CD3-targeting arm to bring tumor cells into close proximity with cytotoxic T cells that mediate cell killing. The molecules comprised multiple bispecific formats bearing different stoichiometries, geometries, and sizes, to enable identification of lead molecules with favorable activities and safety profiles. GPRC5D bispecifics displayed potent T cell-mediated cytotoxicity with picomolar potency. They also exhibited high specificity and bound to only their target, and not the other 6,000 proteins tested using a Membrane Proteome Array (MPA). Furthermore, these molecules showed good developability profiles and minimal cytokine release. GPRC5DxCD3 bispecifics are currently undergoing additional testing for developability and in vivo potency.ConclusionsGPRC5DxCD3 bispecific antibodies hold promise as potent and safe therapeutics for multiple myeloma
Antibodies are important tools for investigating the proteome, but
current methods for producing them have become a rate-limiting step. A primary obstacle in most methods for generating antibodies or
...antibody-like molecules is the requirement for at least microgram quantities of
purified protein. We have developed a technology for producing antibodies using
genetic immunization. Genetic immunization-based antibody
production offers several advantages, including high throughput
and high specificity. Moreover, antibodies produced from genetically immunized
animals are more likely to recognize the native protein. Here we
show that a genetic immunization-based system can be used to efficiently
raise useful antibodies to a wide range of antigens. We accomplished this by
linking the antigen gene to various elements that enhance antigenicity and by
codelivering plasmids encoding genetic adjuvants. Our system, which was tested
by immunizing mice with >130 antigens, has shown a final success rate of
84%.
Facing it Chambers, Ross
1998., 20091110, 1998, 1999, 1998, c1998.
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For a generation or more, literary theorists have used the metaphor of "the death of the author" in considering the observation that to write is to abdicate control over the meanings one's text is ...capable of generating. But in the case of AIDS diaries, the metaphor can be literal. Facing It examines the genre not in classificatory terms but pragmatically, as the site of a social interaction. Through a detailed study of three such diaries, originating respectively in France, the United States, and Australia, Ross Chambers demonstrates that issues concerning the politics of AIDS writing and the ethics of reading are linked by a common concern with the problematics of survivorhood.
BackgroundCTIM-76 is a bispecific Claudin 6 (CLDN6) T-cell engager antibody in preclinical development for treatment of CLDN6-positive cancers, including ovarian, testicular, and non-small cell lung ...cancers. CLDN6 is a tight junction protein differentially expressed on cancer cells with limited, if any expression in normal healthy tissue. Discovery of therapeutic monoclonal antibodies targeting CLDN6 is difficult due to an abundance of closely related family members expressed on healthy tissues and stringent requirements for high specificity. The extracellular region of CLDN6 closely resembles other claudins including CLDN3, CLDN4, and CLDN9 that are expressed in critical organs.MethodsStarting with a panel of high specificity CLDN6 antibodies, we engineered a large set (>50) of CLDN6xCD3 bispecific antibodies using multiple bispecific formats and CD3 arms. These molecules display different geometries and binding stoichiometries which are expected to play a critical role in their in vitro and in vivo potency. In vitro T cell cytotoxicity, cytokine release, and developability studies were used to select the lead candidate now designated as CTIM-76. CTIM-76 is being tested in vivo using xenograft studies in PBMC-engrafted mice.ResultsCTIM-76 demonstrated potent cytotoxic effects on cells expressing CLDN6, and results suggest a wide therapeutic window. Picomolar T cell-dependent cytotoxicity values were obtained with OV90 and OVCAR3 ovarian cancer lines, and studies in K562 cells over-expressing claudin proteins show at least a 500-fold selectivity for CLDN6 over CLDN3, CLDN4, and CLDN9. No CTIM-76 binding was observed against closely related CLDN proteins including CLDN3, CLDN4, and CLDN9, using flow cytometry at CTIM-76 concentrations up to 1 micromolar. In vivo studies with PBMC-engrafted animals using multiple ovarian cancer cell lines are ongoing and preliminary results suggest in vivo potency consistent with the in vitro data. CTIM-76 has entered GMP-manufacturing with productivity levels higher than standard IgG molecules and compatibility with standard purification and formulation. CTIM-76 has also begun PK and GLP-tox studies in cynomolgus monkeys.ConclusionsThe exquisite specificity of CTIM-76 and potent cell-killing effects suggests it has significant potential as a potent and safe therapeutic against CLDN6-positive cancers. Context Therapeutics will be responsible for the clinical development of CTIM-76, and IND filing for this molecule is anticipated to occur in Q1 2024.
Antibodies are quintessential affinity reagents for the investigation and determination of a protein's expression patterns, localization, quantitation, modifications, purification, and functional ...understanding. Antibodies are typically used in techniques such as Western blot, immunohistochemistry (IHC), and enzyme-linked immunosorbent assays (ELISA), among others. The methods employed to generate antibodies can have a profound impact on their success in any of these applications. We raised antibodies against 10 serum proteins using 3 immunization methods: peptide antigens (3 per protein), DNA prime/protein fragment-boost ("DNA immunization"; 3 per protein), and full length protein. Antibodies thus generated were systematically evaluated using several different assay technologies (ELISA, IHC, and Western blot). Antibodies raised against peptides worked predominantly in applications where the target protein was denatured (57% success in Western blot, 66% success in immunohistochemistry), although 37% of the antibodies thus generated did not work in any of these applications. In contrast, antibodies produced by DNA immunization performed well against both denatured and native targets with a high level of success: 93% success in Western blots, 100% success in immunohistochemistry, and 79% success in ELISA. Importantly, success in one assay method was not predictive of success in another. Immunization with full length protein consistently yielded the best results; however, this method is not typically available for new targets, due to the difficulty of generating full length protein. We conclude that DNA immunization strategies which are not encumbered by the limitations of efficacy (peptides) or requirements for full length proteins can be quite successful, particularly when multiple constructs for each protein are used.
Tags are widely used to monitor a protein's expression level, interactions, protein trafficking, and localization. Membrane proteins are often tagged in their extracellular domains to allow ...discrimination between protein in the plasma membrane from that in internal pools. Multipass membrane proteins offer special challenges for inserting a tag since the extracellular regions are often composed of small loops and thus inserting an epitope tag risks perturbing the structure, function, or location of the membrane protein. We have developed a novel tagging system called snorkel where a transmembrane domain followed by a tag is appended to the cytoplasmic C-terminus of the membrane protein. In this way the tag is displayed extracellularly, but structurally separate from the membrane protein. We have tested the snorkel tag system on a diverse panel of membrane proteins including GPCRs and ion channels and demonstrated that it reliably allows for monitoring of the surface expression.
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