Recombinant antibody production evolves into multiple options aimed at yielding reagents suitable for application-specific needs

Marco, Ario de

doi:10.1186/s12934-015-0320-7

Cited by 46 publications

(22 citation statements)

References 127 publications

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“…Antibody genes can be transformed into plants via the transfer DNA of Agrobacterium tumefaciens, which allows for purification of protein from tobacco leaves or rice seeds. 137,148 Alternatively, antibodies can be harvested from mouse or goat milk or chicken eggs using transgenic animals that have human Ig genes in place of the native Ig loci. 137 While these latter systems are expected to produce proteins with human-like glycosylation, plant systems have been engineered to prevent the attachment of potentially immunogenic sugars such as xylose.…”

Section: Other Expression Systemsmentioning

confidence: 99%

Considerations for the Design of Antibody-Based Therapeutics

Goulet

Atkins

2020

Journal of Pharmaceutical Sciences

183

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: Other Expression Systemsmentioning

confidence: 99%

Considerations for the Design of Antibody-Based Therapeutics

Goulet

Atkins

2020

Journal of Pharmaceutical Sciences

183

154

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“…The first class comprises simple organisms as prokaryotic, yeast, fungi or insect cells whose advantages are low costs and good productivity (in the high mg/L range) 5 . Sub-optimal folding and a very different pattern of glycosylation make these systems useful for production of simple non-glycosylated proteins, or for generation of preliminary binding data.…”

Section: Introductionmentioning

confidence: 99%

A long non-coding SINEUP RNA boosts semi-stable production of fully human monoclonal antibodies in HEK293E cells

Sasso

Latino

Froechlich

et al. 2018

mAbs

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Use of monoclonal antibodies is emerging as a highly promising and fast-developing scenario for innovative treatment of viral, autoimmune and tumour diseases. The search for diagnostic and therapeutic antibodies currently depends on in vitro screening approaches, such as phage and yeast display technologies. Antibody production still represents a critical step for preclinical and clinical evaluations. Accordingly, improving production of monoclonal antibodies represents an opportunity, to facilitate downstream target validations. SINEUP RNAs are long non-coding transcripts, possessing the ability to enhance translation of selected mRNAs. We applied SINEUP technology to semi-stable production of monoclonal antibodies in HEK293E cells, which allows for episomal propagation of the expression vectors encoding the heavy and light chains of IgGs. Co-expression of SINEUP RNA with mRNAs encoding heavy and light chains of IgG4s was able to increase the production of different anti-CLDN1 antibodies up to three-fold. Improved production of monoclonal antibodies was achieved both in transiently transfected HEK293E cells and in cellular clones with stable expression of the SINEUP. Compared to antibody preparations obtained under standard conditions, the anti-CLDN1 IgG4s produced in the presence of the SINEUP transcript showed unaltered post-translational modifications, and retained the ability to recognize their target. We thus propose SINEUP technology as a valuable tool to enhance semi-stable antibody production in human cell lines.

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“…Nanobodies are antibody fragments which correspond to the variable region of the heavy‐chain‐only antibodies (VHH) of Camelidae origin. They know a constantly growing interest because of their small mass (14 kDa), high stability and simplicity in being mutated and functionalized . In this study, we show that an anti‐human epidermal growth factor receptor 2 (HER2) nanobody isolated from a pre‐immune library recognized also the canine protein homologue DER2.…”

Section: Introductionmentioning

confidence: 84%

Engineered cross‐reacting nanobodies simplify comparative oncology between humans and dogs

Mazzega

Marco

2017

Vet Comparative Oncology

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Antibodies cross-reacting with homologue antigens in different species would be essential reagents for the development of comparative oncology studies. In comparison with conventional immunoglobulin Gs, recombinant nanobodies (single-domain variable regions of heavy-chain only antibodies of Camelidae origin) can be easily isolated in vitro and engineered into a variety of reagents with optimized characteristics for different research and clinical applications. We recovered an anti-human epidermal growth factor receptor 2 (anti-HER2) nanobody from a naïve llama library by direct panning on whole cells and expressed it fused to Fc and green fluorescent protein. These immunoreagents were assessed by flow cytometry and immunofluorescence with both human and canine cells overexpressing HER2 and its canine homologue dog epidermal growth factor receptor 2. The reported data illustrate the potential of using this class of antibodies in comparative oncology and suggest some development perspectives enabled by in vitro panning of pre-immune nanobody libraries.

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Recombinant antibody production evolves into multiple options aimed at yielding reagents suitable for application-specific needs

Cited by 46 publications

References 127 publications

Considerations for the Design of Antibody-Based Therapeutics

Considerations for the Design of Antibody-Based Therapeutics

A long non-coding SINEUP RNA boosts semi-stable production of fully human monoclonal antibodies in HEK293E cells

Engineered cross‐reacting nanobodies simplify comparative oncology between humans and dogs

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