High-throughput screening for developability during early-stage antibody discovery using self-interaction nanoparticle spectroscopy

Liu, Yuqi; Caffry, Isabelle; Wu, Jugang; Geng, Steven B.; Jain, Tushar; Sun, Tingwan; Reid, Felicia; Cao, Yuan; Estep, Patricia A.; Yu, Yao; Vásquez, Maximiliano; Tessier, Peter M.; Xu, Yingda

doi:10.4161/mabs.27431

Cited by 114 publications

(149 citation statements)

References 44 publications

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“…AC-SINS uses gold nanoparticles pre-coated with anti-human Fc polyclonal antibodies to capture test mAbs. Self-interactions of immobilized mAbs lead to clustering of the gold nanoparticles, which is measured by a shift in absorbance due to changes in their optical properties 21 . The 11 mAbs studied had AC-SINS scores ranging from 0–24 (Table 1).…”

Section: Resultsmentioning

confidence: 99%

Linear pharmacokinetic parameters for monoclonal antibodies are similar within a species and across different pharmacological targets: A comparison between human, cynomolgus monkey and hFcRn Tg32 transgenic mouse using a population-modeling approach

Betts

Keunecke²,

Steeg³

et al. 2018

mAbs

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The linear pharmacokinetics (PK) of therapeutic monoclonal antibodies (mAbs) can be considered a class property with values that are similar to endogenous IgG. Knowledge of these parameters across species could be used to avoid unnecessary in vivo PK studies and to enable early PK predictions and pharmacokinetic/pharmacodynamic (PK/PD) simulations. In this work, population-pharmacokinetic (popPK) modeling was used to determine a single set of ‘typical’ popPK parameters describing the linear PK of mAbs in human, cynomolgus monkey and transgenic mice expressing the human neonatal Fc receptor (hFcRn Tg32), using a rich dataset of 27 mAbs. Non-linear PK was excluded from the datasets and a 2-compartment model was applied to describe mAb disposition. Typical human popPK estimates compared well with data from comparator mAbs with linear PK in the clinic. Outliers with higher than typical clearance were found to have non-specific interactions in an affinity-capture self-interaction nanoparticle spectroscopy assay, offering a potential tool to screen out these mAbs at an early stage. Translational strategies were investigated for prediction of human linear PK of mAbs, including use of typical human popPK parameters and allometric exponents from cynomolgus monkey and Tg32 mouse. Each method gave good prediction of human PK with parameters predicted within 2-fold. These strategies offer alternative options to the use of cynomolgus monkeys for human PK predictions of linear mAbs, based on in silico methods (typical human popPK parameters) or using a rodent species (Tg32 mouse), and call into question the value of completing extensive in vivo preclinical PK to inform linear mAb PK.

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Section: Resultsmentioning

confidence: 99%

Linear pharmacokinetic parameters for monoclonal antibodies are similar within a species and across different pharmacological targets: A comparison between human, cynomolgus monkey and hFcRn Tg32 transgenic mouse using a population-modeling approach

Betts

Keunecke²,

Steeg³

et al. 2018

mAbs

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“…35,36 The H35 is positioned at the VH/ VL interface and is largely buried according to the SASA analysis of the homology model (Table 1). Despite the lack of solvent exposure, mutation of this residue to alanine (H35A) resulted in a significant reduction in mAb-C self-association as evident by the significant decrease in plasmon shift signals measured by AC-SINS (Fig.…”

Section: Affinity-capture Self-interaction Nanoparticle Spectroscopymentioning

confidence: 99%

“…36 Briefly, citrate-stabilized 20 nm gold nanoparticles (Ted Pella Inc., Redding, CA) were combined at a 10:1 ratio with either 0.4 mg/mL ChromePure goat IgG, whole molecule (Jackson ImmunoResearch, West Grove, PA) to yield non-capture nanoparticles, or a 0.4 mg/mL mixture of goat IgG and AffiniPure goat antihuman IgG Fcg fragment specific antibody (Jackson ImmunoResearch), such that 20% of the antibody mixture is goat IgG and 80% is anti-human IgG Fcg, to yield capture nanoparticles. Prior to addition of the nanoparticles, both goat IgG and antihuman IgG Fcg were dialyzed into 20 mM potassium acetate, pH 4.3.…”

Section: Antibody Generationmentioning

confidence: 99%

“…Historically, a number of computational and analytical screening methods have been used to predict aggregation [28][29][30] and high concentration formulation risks. 17,[31][32][33][34][35][36] While these approaches can provide valuable information and help screening of potential lead candidates during discovery and early stage development, they do not necessarily provide insight into the underlying molecular mechanisms involved. With regard to high concentration challenges, the availability of methodologies that can probe native intermolecular interactions at the structural and molecular level can effectively guide corresponding protein engineering and formulation efforts designed to mitigate the development risks.…”

Section: Introductionmentioning

confidence: 99%

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Mitigation of reversible self-association and viscosity in a human IgG1 monoclonal antibody by rational, structure-guided Fv engineering

Geoghegan¹,

Fleming²,

Damschroder³

et al. 2016

mAbs

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Undesired solution behaviors such as reversible self-association (RSA), high viscosity, and liquid-liquid phase separation can introduce substantial challenges during development of monoclonal antibody formulations. Although a global mechanistic understanding of RSA (i.e., native and reversible proteinprotein interactions) is sufficient to develop robust formulation controls, its mitigation via protein engineering requires knowledge of the sites of protein-protein interactions. In the study reported here, we coupled our previous hydrogen-deuterium exchange mass spectrometry findings with structural modeling and in vitro screening to identify the residues responsible for RSA of a model IgG1 monoclonal antibody (mAb-C), and rationally engineered variants with improved solution properties (i.e., reduced RSA and viscosity). Our data show that mutation of either solvent-exposed aromatic residues within the heavy and light chain variable regions or buried residues within the heavy chain/light chain interface can significantly mitigate RSA and viscosity by reducing the IgG's surface hydrophobicity. The engineering strategy described here highlights the utility of integrating complementary experimental and in silico methods to identify mutations that can improve developability, in particular, high concentration solution properties, of candidate therapeutic antibodies.

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“…For monoclonal antibodies, these properties include high-level expression, high solubility, covalent integrity, conformational and colloidal stability, low polyspecificity, and low immunogenicity. The high cost of failing any of these criteria at a late stage in drug development has led to considerable efforts at predicting developability on the basis of sequence motifs and experimentally determined biophysical properties (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15).…”

mentioning

confidence: 99%

Biophysical properties of the clinical-stage antibody landscape

Jain

Sun

Durand

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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470

824

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Antibodies are a highly successful class of biological drugs, with over 50 such molecules approved for therapeutic use and hundreds more currently in clinical development. Improvements in technology for the discovery and optimization of high-potency antibodies have greatly increased the chances for finding binding molecules with desired biological properties; however, achieving drug-like properties at the same time is an additional requirement that is receiving increased attention. In this work, we attempt to quantify the historical limits of acceptability for multiple biophysical metrics of "developability." Amino acid sequences from 137 antibodies in advanced clinical stages, including 48 approved for therapeutic use, were collected and used to construct isotypematched IgG1 antibodies, which were then expressed in mammalian cells. The resulting material for each source antibody was evaluated in a dozen biophysical property assays. The distributions of the observed metrics are used to empirically define boundaries of drug-like behavior that can represent practical guidelines for future antibody drug candidates.monoclonal antibody | developability | biophysical properties | manufacturability | nonspecificity T arget binding is the predominant first concern in development of any drug. However, once a lead molecule attains the desired potency of biological modification, a suite of characteristics termed "developability" assumes critical importance. For monoclonal antibodies, these properties include high-level expression, high solubility, covalent integrity, conformational and colloidal stability, low polyspecificity, and low immunogenicity. The high cost of failing any of these criteria at a late stage in drug development has led to considerable efforts at predicting developability on the basis of sequence motifs and experimentally determined biophysical properties (1-15).In a landmark study of small-molecule drugs over 2,000 molecules with United States Adopted Names (USAN) designations and known to have oral availability were collected and computationally analyzed (16). A simple set of thresholds, encapsulated as the "Lipinski rule of fives," was formulated and has been used by many to prioritize small molecules for entry into clinical development. To date, analogous guiding principles for antibody drugs have not emerged-we therefore endeavor here to do so. By analogy to the Lipinski effort, we first collected the sequences of antibodies that had reached at least phase-2 trials and had USAN or WHO International Nonproprietary Names (INN) designations (137 in total as of the start of this project). As a common basis for comparison of intrinsic variable domain phenotypes we expressed each antibody as the human IgG1 isotype and formulated them in simple Hepes-buffered saline. Each antibody was then subjected to a battery of 12 different biophysical assays in common use for developability assessment.Unexpectedly, for many of the measures the distribution of values was not symmetrically Gaussian, but instead was lon...

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High-throughput screening for developability during early-stage antibody discovery using self-interaction nanoparticle spectroscopy

Abstract: (2014) High-throughput screening for developability during early-stage antibody discovery using self-interaction nanoparticle spectroscopy, mAbs, 6:2, 483-492,

Cited by 114 publications

References 44 publications

Linear pharmacokinetic parameters for monoclonal antibodies are similar within a species and across different pharmacological targets: A comparison between human, cynomolgus monkey and hFcRn Tg32 transgenic mouse using a population-modeling approach

Linear pharmacokinetic parameters for monoclonal antibodies are similar within a species and across different pharmacological targets: A comparison between human, cynomolgus monkey and hFcRn Tg32 transgenic mouse using a population-modeling approach

Mitigation of reversible self-association and viscosity in a human IgG1 monoclonal antibody by rational, structure-guided Fv engineering

Biophysical properties of the clinical-stage antibody landscape

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