Recombinant antibodies for diagnostics and therapy against pathogens and toxins generated by phage display

Kühn, Philipp; Fühner, Viola; Unkauf, Tobias; Moreira, Gustavo Marçal Schmidt Garcia; Frenzel, André; Miethe, Sebastian; Hust, Michael

doi:10.1002/prca.201600002

Cited by 68 publications

(48 citation statements)

References 287 publications

(269 reference statements)

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“…130 Phage display, which uses viruses such as M13 phage, works by fusing the antibody scFv sequence with that of phage surface molecules such as coat protein pIII. 131 The DNA sequence within the plasmid codes for the corresponding surface protein, allowing for phenotypic selection and subsequent genotypic identification. Phage particles that bind antigen strongly are amplified via infection of bacterial hosts such as E. coli.…”

Section: Displaymentioning

confidence: 99%

Considerations for the Design of Antibody-Based Therapeutics

Goulet

Atkins

2020

Journal of Pharmaceutical Sciences

187

154

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Keywords: antibody(s) antibody drug(s) antibody drug conjugate(s) (ADC) physicochemical properties pharmacokinetics monoclonal antibody(s) immunotherapy IgG antibody(s) drug design developability a b s t r a c t Antibody-based proteins have become an important class of biologic therapeutics, due in large part to the stability, specificity, and adaptability of the antibody framework. Indeed, antibodies not only have the inherent ability to bind both antigens and endogenous immune receptors but also have proven extremely amenable to protein engineering. Thus, several derivatives of the monoclonal antibody format, including bispecific antibodies, antibody-drug conjugates, and antibody fragments, have demonstrated efficacy for treating human disease, particularly in the fields of immunology and oncology. Reviewed here are considerations for the design of antibody-based therapeutics, including immunological context, therapeutic mechanisms, and engineering strategies. First, characteristics of antibodies are introduced, with emphasis on structural domains, functionally important receptors, isotypic and allotypic differences, and modifications such as glycosylation. Then, aspects of therapeutic antibody design are discussed, including identification of antigen-specific variable regions, choice of expression system, use of multispecific formats, and design of antibody derivatives based on fragmentation, oligomerization, or conjugation to other functional moieties. Finally, strategies to enhance antibody function through protein engineering are reviewed while highlighting the impact of fundamental biophysical properties on protein developability.

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Section: Displaymentioning

confidence: 99%

Considerations for the Design of Antibody-Based Therapeutics

Goulet

Atkins

2020

Journal of Pharmaceutical Sciences

187

154

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“…So far, several reports demonstrated the successful use of antibody-phage display for the generation of diagnostic antibodies for the detection of pathogenic bacteria [31]. But to our knowledge, this is the first report of a successful selection of a recombinant human antibody against the pathogen L. pneumophila from a naïve human antibody library by antibody-phage display.…”

Section: Discussionmentioning

confidence: 88%

Human Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity

Kühn

Thiem

Steinert

et al. 2019

HAB

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Abstract. Legionella are Gram-negative bacteria that are ubiquitously present in natural and man-made water reservoirs. When humans inhale aerosolized water contaminated with Legionella, alveolar macrophages can be infected, which may lead to a lifethreatening pneumonia called Legionnaires' disease. Due to the universal distribution of Legionella in water and their potential threat to human health, the Legionella concentration in water for human use must be strictly monitored, which is difficult since the standard detection still relies on lengthy cultivation and analysis of bacterial morphology. In this study, an antibody against L. pneumophila has been generated from the naïve human HAL antibody libraries by phage-display for the first time. The panning was performed on whole bacterial cells in order to select antibodies that bind specifically to the cell surface of untreated Legionella. The bacterial cell wall component lipopolysaccharide (LPS) was identified as the target structure. Specific binding to the important pathogenic L. pneumophila strains Corby, Philadelphia-1 and Knoxville was observed, while no binding was detected to seven members of the families Enterobacteriaceae, Pseudomonadaceae or Clostridiaceae. Production of this antibody in the recombinant scFv-Fc format using either a murine or a human Fc part allowed the set-up of a sandwich-ELISA for detection of Legionella cells. The scFv-Fc construct proved to be very stable, even when stored for several weeks at elevated temperatures. A sensitivity limit of 4,000 cells was achieved. The scFv-Fc antibody pair was integrated on a biosensor, demonstrating the specific and fast detection of L. pneumophila on a portable device. With this system, 10,000 Legionella cells were detected within 35 min. Combined with a water filtration/concentration system, this antibody may be developed into a promising reagent for rapid on-site Legionella monitoring.

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“…The extraordinary specificity of antibody binding, with affinities ranging from low nanomolar to picomolar values, allowed for their use in a number of analytical, bioengineering, and biomedical technologies. These affinities have found a myriad of applications in diverse fields and have been the focus of extensive literature reviews (4–6). …”

Section: Introductionmentioning

confidence: 99%

Beyond Antibodies as Binding Partners: The Role of Antibody Mimetics in Bioanalysis

Yang

Dikici

et al. 2017

Annual Rev. Anal. Chem.

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The emergence of novel binding proteins or antibody mimetics capable of binding to ligand analytes in a manner analogous to that of the antigen–antibody interaction has spurred increased interest in the biotechnology and bioanalytical communities. The goal is to produce antibody mimetics designed to outperform antibodies with regard to binding affinities, cellular and tumor penetration, large-scale production, and temperature and pH stability. The generation of antibody mimetics with tailored characteristics involves the identification of a naturally occurring protein scaffold as a template that binds to a desired ligand. This scaffold is then engineered to create a superior binder by first creating a library that is then subjected to a series of selection steps. Antibody mimetics have been successfully used in the development of binding assays for the detection of analytes in biological samples, as well as in separation methods, cancer therapy, targeted drug delivery, and in vivo imaging. This review describes recent advances in the field of antibody mimetics and their applications in bioanalytical chemistry, specifically in diagnostics and other analytical methods.

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Recombinant antibodies for diagnostics and therapy against pathogens and toxins generated by phage display

Cited by 68 publications

References 287 publications

Considerations for the Design of Antibody-Based Therapeutics

Considerations for the Design of Antibody-Based Therapeutics

Human Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity

Beyond Antibodies as Binding Partners: The Role of Antibody Mimetics in Bioanalysis

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