Liquid Formulations for Stabilizing IgMs During Physical Stress and Long-Term Storage

Mueller, Monika; Loh, Maybelle Q. T.; Tscheließnig, Rupert; Tee, Doris Hui Ying; E, Tan; Bardor, Muriel; Jungbauer, Alois

doi:10.1007/s11095-012-0914-2

Cited by 21 publications

(19 citation statements)

References 44 publications

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“…The protein form factor is the Fourier transform of the pair correlation of the crystallographic data of cutinase. The pair correlation was computed by taking into account all sites as possible sources of scattering (20)(21)(22).…”

Section: Methodsmentioning

confidence: 99%

Enhanced Cutinase-Catalyzed Hydrolysis of Polyethylene Terephthalate by Covalent Fusion to Hydrophobins

Ribitsch

Acero

Przylucka

et al. 2015

Appl Environ Microbiol

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164

102

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g Cutinases have shown potential for hydrolysis of the recalcitrant synthetic polymer polyethylene terephthalate (PET). We have shown previously that the rate of this hydrolysis can be enhanced by the addition of hydrophobins, small fungal proteins that can alter the physicochemical properties of surfaces. Here we have investigated whether the PET-hydrolyzing activity of a bacterial cutinase from Thermobifida cellulosilytica (Thc_Cut1) would be further enhanced by fusion to one of three Trichoderma hydrophobins, i.e., the class II hydrophobins HFB4 and HFB7 and the pseudo-class I hydrophobin HFB9b. The fusion enzymes exhibited decreased k cat values on soluble substrates (p-nitrophenyl acetate and p-nitrophenyl butyrate) and strongly decreased the hydrophilicity of glass but caused only small changes in the hydrophobicity of PET. When the enzyme was fused to HFB4 or HFB7, the hydrolysis of PET was enhanced >16-fold over the level with the free enzyme, while a mixture of the enzyme and the hydrophobins led only to a 4-fold increase at most. Fusion with the non-class II hydrophobin HFB9b did not increase the rate of hydrolysis over that of the enzyme-hydrophobin mixture, but HFB9b performed best when PET was preincubated with the hydrophobins before enzyme treatment. The pattern of hydrolysis by the fusion enzymes differed from that of Thc_Cut1 as the concentration of the product mono(2-hydroxyethyl) terephthalate relative to that of the main product, terephthalic acid, increased. Small-angle X-ray scattering (SAXS) analysis revealed an increased scattering contrast of the fusion proteins over that of the free proteins, suggesting a change in conformation or enhanced protein aggregation. Our data show that the level of hydrolysis of PET by cutinase can be significantly increased by fusion to hydrophobins. The data further suggest that this likely involves binding of the hydrophobins to the cutinase and changes in the conformation of its active center. P olyethylene terephthalate (PET) is the best-known and most widespread synthetic polyester in the world. It is used for foils and bottles as well as for fibers for the textile industry (1). Recycling of PET and modification of its properties for different applications by traditional procedures involve harsh chemical and physicochemical treatments (2, 3). Enzymatic modification, particularly by cutinases, has been recognized as a powerful alternative in the past decade (4, 5) and, besides offering new avenues for PET recycling, has the additional advantage of creating a modified PET with increased dyeing efficacy and improved binding to polyvinyl chloride without altering the polymer's bulk properties (4, 5).On the other hand, enzymatic hydrolysis of PET has the inherent disadvantage of occurring at a very low rate (5, 6). The reasons for this are not yet clearly understood. The access of the active center of the cutinase to the insoluble substrate is apparently one of the rate-limiting points, because enlarging the areas around the active sites of cutinases from Fus...

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Section: Methodsmentioning

confidence: 99%

Enhanced Cutinase-Catalyzed Hydrolysis of Polyethylene Terephthalate by Covalent Fusion to Hydrophobins

Ribitsch

Acero

Przylucka

et al. 2015

Appl Environ Microbiol

Self Cite

164

102

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“…Another study examined the stability of eight mouse mAbs and concluded that DSC was predictive of accelerated and long term storage for six out eight of the mAbs studied 39 . Cases in which conformational stability did not clearly predict physical stability of mAbs have also been reported.…”

Section: Introductionmentioning

confidence: 99%

Examination of Thermal Unfolding and Aggregation Profiles of a Series of Developable Therapeutic Monoclonal Antibodies

et al. 2015

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Screening for pharmaceutically viable stability from measurements of thermally induced protein unfolding and short-term accelerated stress underpins much molecule design, selection, and formulation in the pharmaceutical biotechnology industry. However, the interrelationships among intrinsic protein conformational stability, thermal denaturation, and pharmaceutical stability are complex. There are few publications in which predictions from thermal unfolding-based screening methods are examined together with pharmaceutically relevant long-term storage stability performance. We have studied eight developable therapeutic IgG molecules under solution conditions optimized for large-scale commercial production and delivery. Thermal unfolding profiles were characterized by differential scanning calorimetry (DSC) and intrinsic fluorescence recorded simultaneously with static light scattering (SLS). These molecules exhibit a variety of thermal unfolding profiles under common reference buffer conditions and under individually optimized formulation conditions. Aggregation profiles by SE-HPLC and bioactivity upon long-term storage at 5, 25, and 40 °C establish that IgG molecules possessing a relatively wide range of conformational stabilities and thermal unfolding profiles can be formulated to achieve pharmaceutically stable drug products. Our data suggest that a formulation design strategy that increases the thermal unfolding temperature of the Fab transition may be a better general approach to improving pharmaceutical storage stability than one focused on increasing Tonset or Tm of the first unfolding transition.

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“…Preclinical and clinical studies have underscored the efficacy against various infectious agents of polyclonal IgM-enriched preparations administered by the systemic route (22)(23)(24)(25)(26). Similar to SIgA, SIgM can be seen as a valid candidate immunoglobulin for mucosal application, given its ability to bind antigens with strong avidity, its potential to ensure long term protection (27), its capacity to survive low pH conditions (28), as well as its resistance to proteases (29). We have demonstrated recently that human plasma can serve as a source of polyreactive, polymeric IgA (pIgA) and IgM to generate secretory-like IgA and IgM Abs, the natural molecular form found in secretions.…”

mentioning

confidence: 99%

Reconstituted Human Polyclonal Plasma-derived Secretory-like IgM and IgA Maintain the Barrier Function of Epithelial Cells Infected with an Enteropathogen

Longet

Vonarburg

Lötscher

et al. 2014

Journal of Biological Chemistry

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Liquid Formulations for Stabilizing IgMs During Physical Stress and Long-Term Storage

Abstract: A long-term stabilizing formulation for 8 IgMs was found comprising 20% sorbitol and 1 M glycine at pH 5.0-5.5 which may have broad utility for other IgMs. Formulation development using DSC and accelerated storage was evaluated in this study and may be used for other proteins.

Cited by 21 publications

References 44 publications

Enhanced Cutinase-Catalyzed Hydrolysis of Polyethylene Terephthalate by Covalent Fusion to Hydrophobins

Enhanced Cutinase-Catalyzed Hydrolysis of Polyethylene Terephthalate by Covalent Fusion to Hydrophobins

Examination of Thermal Unfolding and Aggregation Profiles of a Series of Developable Therapeutic Monoclonal Antibodies

Reconstituted Human Polyclonal Plasma-derived Secretory-like IgM and IgA Maintain the Barrier Function of Epithelial Cells Infected with an Enteropathogen

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