An automated immunoassay for early specificity profiling of antibodies

Frese, Katrin; Eisenmann, Meike; Ostendorp, Ralf; Brocks, Bodo; Pabst, Stefan

doi:10.4161/mabs.23539

Cited by 13 publications

(17 citation statements)

References 26 publications

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“…However, during early stage development, it is important to eliminate antibodies with off-target reactivity. This may be done using assays assessing binding to panels of non-cognate proteins 60 – 62 . Interestingly, it was recently shown that this may simply be done by screening the antibodies towards recombinant chaperone proteins that correlated with clearance in mice 63 .…”

Section: Discussionmentioning

confidence: 99%

A human endothelial cell-based recycling assay for screening of FcRn targeted molecules

et al. 2018

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Albumin and IgG have remarkably long serum half-lives due to pH-dependent FcRn-mediated cellular recycling that rescues both ligands from intracellular degradation. Furthermore, increase in half-lives of IgG and albumin-based therapeutics has the potential to improve their efficacies, but there is a great need for robust methods for screening of relative FcRn-dependent recycling ability. Here, we report on a novel human endothelial cell-based recycling assay (HERA) that can be used for such pre-clinical screening. In HERA, rescue from degradation depends on FcRn, and engineered ligands are recycled in a manner that correlates with their half-lives in human FcRn transgenic mice. Thus, HERA is a novel cellular assay that can be used to predict how FcRn-binding proteins are rescued from intracellular degradation.

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Section: Discussionmentioning

confidence: 99%

A human endothelial cell-based recycling assay for screening of FcRn targeted molecules

et al. 2018

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“…The need for well-characterized, defined reagents has led to the use of Hsp90 and insulin as single-component specificity reagents that also correlate to antibody clearance rates in mice [ 196 , 199 ]. Previous specificity assays, employing microarray technologies or microtiter plate assays, afford much lower throughput (tens to hundreds) relative to FACS-based approaches that can process millions of variants [ 200 ].…”

Section: Polyspecificty and In Vivo Propertiesmentioning

confidence: 99%

Toward Drug-Like Multispecific Antibodies by Design

Sawant

Streu

et al. 2020

IJMS

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The success of antibody therapeutics is strongly influenced by their multifunctional nature that couples antigen recognition mediated by their variable regions with effector functions and half-life extension mediated by a subset of their constant regions. Nevertheless, the monospecific IgG format is not optimal for many therapeutic applications, and this has led to the design of a vast number of unique multispecific antibody formats that enable targeting of multiple antigens or multiple epitopes on the same antigen. Despite the diversity of these formats, a common challenge in generating multispecific antibodies is that they display suboptimal physical and chemical properties relative to conventional IgGs and are more difficult to develop into therapeutics. Here we review advances in the design and engineering of multispecific antibodies with drug-like properties, including favorable stability, solubility, viscosity, specificity and pharmacokinetic properties. We also highlight emerging experimental and computational methods for improving the next generation of multispecific antibodies, as well as their constituent antibody fragments, with natural IgG-like properties. Finally, we identify several outstanding challenges that need to be addressed to increase the success of multispecific antibodies in the clinic.

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“…43 Frese et al used Protein Panel Profiling (3P) to measure polyreactivity of commercially available therapeutic proteins for a panel of 32 test human proteins that were selected based on theoretical considerations and results from previous experiments. 44 They demonstrated high specificity for all tested therapeutic proteins, including 4 monoclonal antibodies (adalimumab (Humira Ò ), bevacizumab (Avastin Ò ), rituximab (Rituxan Ò ), ustekinumab (Stelara Ò )), with no significant off-target binding. Overall these results support the use of protein chip assay as an effective vitro screening tool in drug discovery.…”

Section: Polyspecificity Binding Assaymentioning

confidence: 99%

Pharmacokinetic de-risking tools for selection of monoclonal antibody lead candidates

Dostálek

Prueksaritanont

Kelley³

2017

mAbs

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Pharmacokinetic studies play an important role in all stages of drug discovery and development. Recent advancements in the tools for discovery and optimization of therapeutic proteins have created an abundance of candidates that may fulfill target product profile criteria. Implementing a set of in silico, small scale in vitro and in vivo tools can help to identify a clinical lead molecule with promising properties at the early stages of drug discovery, thus reducing the labor and cost in advancing multiple candidates toward clinical development. In this review, we describe tools that should be considered during drug discovery, and discuss approaches that could be included in the pharmacokinetic screening part of the lead candidate generation process to de-risk unexpected pharmacokinetic behaviors of Fc-based therapeutic proteins, with an emphasis on monoclonal antibodies.

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An automated immunoassay for early specificity profiling of antibodies

Cited by 13 publications

References 26 publications

A human endothelial cell-based recycling assay for screening of FcRn targeted molecules

A human endothelial cell-based recycling assay for screening of FcRn targeted molecules

Toward Drug-Like Multispecific Antibodies by Design

Pharmacokinetic de-risking tools for selection of monoclonal antibody lead candidates

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