Interactions and phase behavior of a monoclonal antibody

Lewus, Rachael A.; Darcy, Patricia; Lenhoff, Abraham M.; Sandler, Stanley I.

doi:10.1002/btpr.536

Cited by 66 publications

(119 citation statements)

References 38 publications

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“…While LLPS in solutions of globular proteins is well documented (16,(18)(19)(20)(21), it is often preempted by aggregation or crystallization. Recently, reports have appeared of such LLPS in solutions of antibodies (5)(6)(7)(8)(9)22). Antibodies can be present in blood at relatively high concentrations.…”

Section: Discussionmentioning

confidence: 99%

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Phase separation in solutions of monoclonal antibodies and the effect of human serum albumin

Wang

Lomakin²,

Latypov³

et al. 2011

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

106

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We report the observation of liquid-liquid phase separation in a solution of human monoclonal antibody, IgG2, and the effects of human serum albumin, a major blood protein, on this phase separation. We find a significant reduction of phase separation temperature in the presence of albumin, and a preferential partitioning of the albumin into the antibody-rich phase. We provide a general thermodynamic analysis of the antibody-albumin mixture phase diagram and relate its features to the magnitude of the effective interprotein interactions. Our analysis suggests that additives (HSA in this report), which have moderate attraction with antibody molecules, may be used to forestall undesirable proetin condensation in antibody solutions. Our findings are relevant to understanding the stability of pharmaceutical solutions of antibodies and the mechanisms of cryoglobulinemia.ntibodies are widely used in research and biotechnology, as well as in medical and pharmaceutical applications. In some cases, concentrated solutions of specific antibodies are required. In particular, monoclonal antibodies (MAb) have become a major category of drugs in the treatment of a variety of diseases (1). In some drug delivery routes e.g., subcutaneous administration, formulations with concentrated antibody solutions are required to achieve therapeutic dosing (2).The physiological functions of antibodies are mostly determined by antibody-antigen and antibody-receptor specific interactions. However, the nonspecific interactions between antibodies (i.e., self-association) in the concentrated antibody solutions can also affect their functions. Nonspecific attractive interactions can cause various forms of condensation, including liquid-liquid phase separation, aggregation, and crystallization. Upon such condensation, antibodies lose their solubility, and may lose their biological activity. Particularly, in pharmaceutical industry, these processes impact storage stability and safety of protein therapeutics thus impeding drug development (3). For example, immunogenicity of some biologics has been attributed to formation of protein aggregates (4). The mechanisms of protein condensation are complex and depend on protein concentration, buffer composition, temperature, etc. Clearly, the factors which affect protein condensation throughout the shelf-life must be understood and controlled to ensure biotherapeutic effectiveness.One important condensation process is liquid-liquid phase separation (LLPS). In LLPS, a homogeneous protein solution spontaneously separates into two coexisting phases with different protein concentrations. This phenomenon takes place upon changing the temperature or other solution conditions, and is reversible. In contrast to aggregation or crystallization, LLPS, while highly sensitive to the average "net" attractive interaction between proteins, are much less sensitive to the distribution pattern and the nature of the "local" interactions on the protein surface. As a result, LLPS exhibits universal features applicable to a variety...

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Section: Discussionmentioning

confidence: 99%

“…Recently, LLPS of several pharmaceutical antibodies have been reported (5)(6)(7)(8)(9). There are five isotypes of mammalian antibodies with distinct Fc regions, including IgA, IgD, IgE, IgG, and IgM.…”

mentioning

confidence: 99%

Phase separation in solutions of monoclonal antibodies and the effect of human serum albumin

Wang

Lomakin²,

Latypov³

et al. 2011

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

106

View full text Add to dashboard Cite

show abstract

“…Protein aggregation can be driven by vastly different mechanisms or processes and eventually leads to crystallization, precipitation, gelation, or liquid-liquid phase separation [8][9][10]. Random formation of amorphous, visible aggregates defines protein precipitation, whereas ordered three-dimensional lattices define a crystallite as a particular form of protein precipitate.…”

Section: Introductionmentioning

confidence: 99%

Colloidal interactions between monoclonal antibodies in aqueous solutions

Arzenšek

Kuzman²,

Podgornik

2012

Journal of Colloid and Interface Science

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“…Although IgGs are typically quite soluble at physiological conditions, sometimes they can become insoluble. In fact, recent studies of protein condensation have been published both for recombinant pharmaceutical IgGs and monoclonal IgGs [10][11][12][13][14][15][16][17]. A detailed discussion of the importance of IgGs in physiological and pharmaceutical situations is given by Wang et al [6], as well as Nezlin [18].…”

Section: Introductionmentioning

confidence: 99%

Impact of surface interactions on the phase behavior of Y-shaped molecules

et al. 2015

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In recent years the statistical mechanics of non-spherical molecules, such as polypeptide chains and protein molecules, has garnered considerable attention as their phase behavior has important scientific and health implications. One example is provided by immunoglobulin, which has a "Y"-shape. In this work, we determine the phase diagram of Y-shaped molecules on a hexagonal lattice through Monte Carlo Grand Canonical ensemble simulation, using histogram reweighting, multicanonical sampling, and finite-size scaling. We show that (as expected) this model is a member of the Ising universality class. For low temperatures, we implemented multicanonical sampling to induce faster phase transitions in the simulation. By studying several system sizes, we use finite-size scaling to determine the two phase coexistence curve, including the bulk critical temperature, critical chemical potential, and critical density.

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Interactions and phase behavior of a monoclonal antibody

Cited by 66 publications

References 38 publications

Phase separation in solutions of monoclonal antibodies and the effect of human serum albumin

Phase separation in solutions of monoclonal antibodies and the effect of human serum albumin

Colloidal interactions between monoclonal antibodies in aqueous solutions

Impact of surface interactions on the phase behavior of Y-shaped molecules

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