Antibody Response to Aggregated Human Interferon Alpha2b in Wild-type and Transgenic Immune Tolerant Mice Depends on Type and Level of Aggregation

Hermeling, Suzanne; Schellekens, Huub; Maas, Coen; Gebbink, Martijn F.B.G.; Crommelin, Daan J.A.; Jiskoot, Wim

doi:10.1002/jps.20599

Cited by 173 publications

(138 citation statements)

References 28 publications

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“…16 Such irreversible aggregates can decrease protein activity and stability, and may elicit adverse immunogenic reactions in patients. [17][18][19] In addition, parenteral administration of highly viscous liquids requires thicker gauge needles that may cause more painful injections. 2 Previous work has shown that reversible self-association (RSA) between different IgG1 mAbs can result from different binding interfaces, despite high sequence similarity between the mAbs.…”

Section: Introductionmentioning

confidence: 99%

Charge-mediated Fab-Fc interactions in an IgG1 antibody induce reversible self-association, cluster formation, and elevated viscosity

Arora

Esfandiary³

et al. 2016

mAbs

103

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Concentration-dependent reversible self-association (RSA) of monoclonal antibodies (mAbs) poses a challenge to their pharmaceutical development as viable candidates for subcutaneous delivery. While the role of the antigen-binding fragment (Fab) in initiating RSA is well-established, little evidence supports the involvement of the crystallizable fragment (Fc). In this report, a variety of biophysical tools, including hydrogen exchange mass spectrometry, are used to elucidate the protein interface of such non-covalent protein-protein interactions. Using dynamic and static light scattering combined with viscosity measurements, we find that an IgG1 mAb (mAb-J) undergoes RSA primarily through electrostatic interactions and forms a monomer-dimer-tetramer equilibrium. We provide the first direct experimental mapping of the interface formed between the Fab and Fc domains of an antibody at high protein concentrations. Charge distribution heterogeneity between the positively charged interface spanning complementarity-determining regions CDR3H and CDR2L in the Fab and a negatively charged region in C H 3/Fc domain mediates the RSA of mAb-J. When arginine and NaCl are added, they disrupt RSA of mAb-J and decrease the solution viscosity. Fab-Fc domain interactions between mAb monomers may promote the formation of large transient antibody complexes that ultimately cause increases in solution viscosity. Our findings illustrate how limited specific arrangements of amino-acid residues can cause mAbs to undergo RSA at high protein concentrations and how conserved regions in the Fc portion of the antibody can also play an important role in initiating weak and transient protein-protein interactions.

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Section: Introductionmentioning

confidence: 99%

Charge-mediated Fab-Fc interactions in an IgG1 antibody induce reversible self-association, cluster formation, and elevated viscosity

Arora

Esfandiary³

et al. 2016

mAbs

103

View full text Add to dashboard Cite

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“…16) In addition, Hermeling et al reported that aggregates of recombinant human interferon a2b (rhIFNa2b), which were induced by metalcatalyzed oxidation, exhibited immunogenic reactions in transgenic mice that had been conferred tolerant to rhIFNa2b. 17) Although several case reports have noted the immunogenic reactions of individual proteins, the mechanisms by which therapeutic proteins express immunogenicity remain unclear. 8,9) It is thus critical to minimize the deterioration of proteins to the maximum possible extent to prevent the risk of unexpected immunogenic reactions.…”

mentioning

confidence: 99%

Influence of pH on Heat-Induced Aggregation and Degradation of Therapeutic Monoclonal Antibodies

Ishikawa

Ito

Endo

et al. 2010

Biological & Pharmaceutical Bulletin

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“…Such aggregation leads to fouling of aseptic filters during their manufacture, and may lead to immunogenic reactions in patients. [4][5][6] In the presence of appropriate concentrations of salts that suppress ionic interactions, hydrophobic interactions between the hydrophobic surfaces of protein molecules are the major cause of protein aggregation. Protein denaturation exposes hydrophobic residues buried in the protein core and exacerbates protein aggregation.…”

mentioning

confidence: 99%

Prevention of Stirring-Induced Microparticle Formation in Monoclonal Antibody Solutions

Ishikawa

Kobayashi

Osawa

et al. 2010

Biological & Pharmaceutical Bulletin

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Monoclonal antibodies are being widely used for the treatment of various diseases. Microparticle formation in high-concentration protein solutions is a major problem during the manufacture of therapeutic monoclonal antibodies, because aggregation leads to fouling of aseptic filters and may lead to an immunogenic reaction in patients. We found that stirring using a traditional bottom-magnetic type stirrer results in extensive and sustained formation of 500-nm diameter protein microparticles arising from shear stress on protein molecules. The antibody solution stirred for only 5 min using this type of stirrer exhibited significant fouling of aseptic filter membranes. In contrast, a top-entering type stirrer did not lead to the formation of microparticles, and the solution did not exhibit membrane fouling even after 30 min of stirring. We conclude that a top-entering type stirrer is more suited for the manufacture of concentrated therapeutic monoclonal antibody solutions.

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Antibody Response to Aggregated Human Interferon Alpha2b in Wild-type and Transgenic Immune Tolerant Mice Depends on Type and Level of Aggregation

Cited by 173 publications

References 28 publications

Charge-mediated Fab-Fc interactions in an IgG1 antibody induce reversible self-association, cluster formation, and elevated viscosity

Charge-mediated Fab-Fc interactions in an IgG1 antibody induce reversible self-association, cluster formation, and elevated viscosity

Influence of pH on Heat-Induced Aggregation and Degradation of Therapeutic Monoclonal Antibodies

Prevention of Stirring-Induced Microparticle Formation in Monoclonal Antibody Solutions

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