High-Throughput Screening of Excipients Intended to Prevent Antigen Aggregation at Air-Liquid Interface

Dasnoy, Sébastien; Dezutter, Nancy; Lemoine, D.; Bras, Vivien Le; Préat, Véronique

doi:10.1007/s11095-011-0393-x

Cited by 24 publications

(32 citation statements)

References 43 publications

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“…Other excipients used to prevent adsorption at the air–liquid interface include cyclo-dextrin derivatives (Serno et al, 2010) and amino acids such as arginine, lysine, and glutamic acid (Dasnoy et al, 2011). Carbohydrates such as sucrose, dextrose, and trehalose have also been used as stabilizing excipients (Katakam & Banga, 1995; Kreilgaard et al, 1998a).…”

Section: Methods To Reduce Aggregationmentioning

confidence: 99%

“…Dasnoy et al (2011) developed a HTS stress test for studying aggregates at the air–liquid interface, allowing the evaluation of a large number of excipients for prevention of aggregation. Considerable variation (for example, in size and structure) is observed in aggregates formed during bioprocessing.…”

Section: Methods To Reduce Aggregationmentioning

confidence: 99%

“…Such interfaces are encountered at various points in the life of a biotherapeutic product (Dasnoy et al, 2011). Different biotherapeutic products have also been reported to aggregate at the air–liquid interface; here proteins can act as a detergent, migrating to the air–liquid interface, and this stress can cause partial unfolding.…”

Section: Protein Aggregationmentioning

confidence: 99%

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Immunogenicity of therapeutic proteins: Influence of aggregation

Ratanji

Derrick

Dearman

et al. 2013

Journal of Immunotoxicology

492

351

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The elicitation of anti-drug antibodies (ADA) against biotherapeutics can have detrimental effects on drug safety, efficacy, and pharmacokinetics. The immunogenicity of biotherapeutics is, therefore, an important issue. There is evidence that protein aggregation can result in enhanced immunogenicity; however, the precise immunological and biochemical mechanisms responsible are poorly defined. In the context of biotherapeutic drug development and safety assessment, understanding the mechanisms underlying aggregate immunogenicity is of considerable interest. This review provides an overview of the phenomenon of protein aggregation, the production of unwanted aggregates during bioprocessing, and how the immune response to aggregated protein differs from that provoked by non-aggregated protein. Of particular interest is the nature of the interaction of aggregates with the immune system and how subsequent ADA responses are induced. Pathways considered here include ‘classical’ activation of the immune system involving antigen presenting cells and, alternatively, the breakdown of B-cell tolerance. Additionally, methods available to screen for aggregation and immunogenicity will be described. With an increased understanding of aggregation-enhanced immune responses, it may be possible to develop improved manufacturing and screening processes to avoid, or at least reduce, the problems associated with ADA.

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Section: Methods To Reduce Aggregationmentioning

confidence: 99%

Section: Methods To Reduce Aggregationmentioning

confidence: 99%

Section: Protein Aggregationmentioning

confidence: 99%

See 1 more Smart Citation

Immunogenicity of therapeutic proteins: Influence of aggregation

Ratanji

Derrick

Dearman

et al. 2013

Journal of Immunotoxicology

492

351

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show abstract

“…In medicine, preventing aggregation can improve the consistency and bioavailability of therapeutics, facilitate production and storage, safeguard drug activity, and curb immunogenicity [10]. Methods for inhibiting protein aggregation have been highly sought to improve biopharmaceuticals, from rational design to introduction of excipients [11], [12], [13]. For example, human calcitonin is a small peptide hormone required for calcium regulation and bone formation that is prone to forming amyloid fibrils.…”

Section: Introductionmentioning

confidence: 99%

Alternative Computational Protocols for Supercharging Protein Surfaces for Reversible Unfolding and Retention of Stability

et al. 2013

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Reengineering protein surfaces to exhibit high net charge, referred to as “supercharging”, can improve reversibility of unfolding by preventing aggregation of partially unfolded states. Incorporation of charged side chains should be optimized while considering structural and energetic consequences, as numerous mutations and accumulation of like-charges can also destabilize the native state. A previously demonstrated approach deterministically mutates flexible polar residues (amino acids DERKNQ) with the fewest average neighboring atoms per side chain atom (AvNAPSA). Our approach uses Rosetta-based energy calculations to choose the surface mutations. Both protocols are available for use through the ROSIE web server. The automated Rosetta and AvNAPSA approaches for supercharging choose dissimilar mutations, raising an interesting division in surface charging strategy. Rosetta-supercharged variants of GFP (RscG) ranging from −11 to −61 and +7 to +58 were experimentally tested, and for comparison, we re-tested the previously developed AvNAPSA-supercharged variants of GFP (AscG) with +36 and −30 net charge. Mid-charge variants demonstrated ∼3-fold improvement in refolding with retention of stability. However, as we pushed to higher net charges, expression and soluble yield decreased, indicating that net charge or mutational load may be limiting factors. Interestingly, the two different approaches resulted in GFP variants with similar refolding properties. Our results show that there are multiple sets of residues that can be mutated to successfully supercharge a protein, and combining alternative supercharge protocols with experimental testing can be an effective approach for charge-based improvement to refolding.

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“…In addition to increased quiescent storage stability, both lyophilized and frozen products are less sensitive to mechanical stress, for example, as experienced during transport. In the case of liquid formulations, mechanical stress during transport (e.g., shaking) might result in significant changes of critical quality attributes, necessitating extensive transport studies 4,5. A further advantage of frozen storage is a significant reduction of microbial growth, which is especially relevant in the case of bulk drug substance storage.…”

Section: Introductionmentioning

confidence: 99%

Correction: Betaine for Nonalcoholic Fatty Liver Disease: Results of A Randomized Placebo-Controlled Trial

Abdelmalek¹,

Sanderson²,

Angulo³

et al. 2010

Hepatology

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High-Throughput Screening of Excipients Intended to Prevent Antigen Aggregation at Air-Liquid Interface

Abstract: A high-throughput screening approach can be followed for evaluating the performance of excipients against aggregation of a protein antigen at air-liquid interface.

Cited by 24 publications

References 43 publications

Immunogenicity of therapeutic proteins: Influence of aggregation

Immunogenicity of therapeutic proteins: Influence of aggregation

Alternative Computational Protocols for Supercharging Protein Surfaces for Reversible Unfolding and Retention of Stability

Correction: Betaine for Nonalcoholic Fatty Liver Disease: Results of A Randomized Placebo-Controlled Trial

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