Trehalose Preserves the Integrity of Lyophilized Phycoerythrin–AntiHuman CD8 Antibody Conjugates and Enhances their Thermal Stability in Flow Cytometric Assays

Selva, Corrado; Malferrari, Marco; Ballardini, Rossana; Ventola, Alfredo; Francia, Francesco; Venturoli, Giovanni

doi:10.1002/jps.23398

Cited by 12 publications

(11 citation statements)

References 48 publications

(55 reference statements)

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“…Among disaccharides, trehalose appears to be the most effective in stabilizing biostructures [14]. Its potentialities in providing long-term, room temperature stability to vaccines [15] (i.e., biomedical preparations that improve immunity to a particular disease) and immunoconjugates (i.e., antibody-fluorescent protein conjugates for diagnostic purposes [16], as well as antibody-drug conjugates that have only recently entered the arsenal of anti-cancer drugs [17]) have been pointed out in the literature. This property of trehalose can also be harnessed in the stabilization of liposomes, isolated proteins and in the preservation of red blood cells, but the underlying mechanism of in vitro bioprotection is not yet fully understood.…”

Section: The Anhydrobiotic State and Suggested Mechanisms Of Biostabimentioning

confidence: 99%

The Magic of Disaccharide Glass Matrices for Protein Function as Decoded by High-Field EPR and FTIR Spectroscopy

et al. 2015

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The structural and dynamical interaction of proteins with their microenvironment in disordered matrices plays a decisive role for their function; EPR spectroscopy is a powerful tool for shading light onto the molecular mechanisms of this protein-matrix interplay. To clarify the molecular mechanisms of disaccharide bioprotection, we studied the structure and dynamics of spin-labeled systems and photosynthetic reaction centers (RCs) in sucrose and trehalose matrices at different hydration levels by means of cw and pulse high-field 95 GHz (W-band) EPR as well as by FTIR. In this minireview, we summarize and discuss EPR and FTIR experiments showing that the anhydrobiotic state of the RC-trehalose system (1) is not the result of matrix-induced changes of the local structure of the charge-separated radical-pair cofactors, P Áþ 865 and Q ÁÀ A , and (2) is not the result of changes of local dynamics and local hydrogen bonding of Q A in its binding pocket. Rather, the extreme impairment of RC dynamics caused by incorporation into the dehydrated trehalose matrix, which also protects it against thermal denaturation, originates in the high rigidity, already at room temperature, of the dry trehalose glass matrix coating the RC protein surface. This surface hydrogen-bonding scaffold shifts the correlation time of thermal conformational fluctuations into the non-biological time domain. Another intriguing aspect of disaccharide bioprotection is the superior The authors dedicate this work to Giovanni Giacometti on the occasion of his 85th birthday. AppliedMagnetic Resonance efficiency of trehalose versus sucrose matrices in stabilizing the anhydrobiotic state of proteins. To clarify the molecular basis of this specificity, glassy trehalose-water and sucrose-water binary systems, incorporating a nitroxide radical as spin probe, have been studied by high-field W-band EPR spectroscopy at different water contents. Analysis of the EPR spectra revealed a different structural and dynamical organization in the sucrose and trehalose matrix, only the trehalose being homogeneous in terms of residual water and nitroxide distribution.

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Section: The Anhydrobiotic State and Suggested Mechanisms Of Biostabimentioning

confidence: 99%

The Magic of Disaccharide Glass Matrices for Protein Function as Decoded by High-Field EPR and FTIR Spectroscopy

et al. 2015

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“…11 Among disaccharides, trehalose appears to be the most effective in stabilizing biostructures. 6 Its potentialities in providing long-term, room temperature stability to vaccines 12 and immunoconjugates 13 have been pointed out in the literature.…”

Section: Introductionmentioning

confidence: 99%

“…49 Molecular dynamics simulations of trehalose-, maltose-and sucrose-water solutions revealed that trehalose is able to form larger clusters. 46 A comparative study of cross-polarization-magic-angle-spinning (CPMAS) 13 C NMR performed on trehalose, sucrose and lactose in the amorphous glassy state, paying particular attention to the glycosidic linkage C-atoms, has provided evidence of an enhanced trehalose molecular flexibility, suggesting that the conformational space explored by trehalose molecules in the amorphous phase is larger than that estimated for the other two sugars. 50 Finally, by using positron annihilation lifetime spectroscopy in trehalose matrices, 5 it was found that in the amorphous phase hydration increases the average intermolecular hole size, while in the crystalline trehalose dihydrate water is organized as a onedimensional fluid in channels of fixed diameters, which allow water diffusion in and out of the crystallites, thereby acting as both a sink and a source of water at low moisture.…”

Section: Introductionmentioning

confidence: 99%

Structural and dynamical characteristics of trehalose and sucrose matrices at different hydration levels as probed by FTIR and high-field EPR

Malferrari

Nalepa

Venturoli

et al. 2014

Phys. Chem. Chem. Phys.

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Some organisms can survive complete dehydration and high temperatures by adopting an anhydrobiotic state in which the intracellular medium contains large amounts of disaccharides, particularly trehalose and sucrose. Trehalose is most effective also in protecting isolated in vitro biostructures. In an attempt to clarify the molecular mechanisms of disaccharide bioprotection, we compared the structure and dynamics of sucrose and trehalose matrices at different hydration levels by means of high-field W-band EPR and FTIR spectroscopy. The hydration state of the samples was characterized by FTIR spectroscopy and the structural organization was probed by EPR using a nitroxide radical dissolved in the respective matrices. Analysis of the EPR spectra showed that the structure and dynamics of the dehydrated matrices as well as their evolution upon re-hydration differ substantially between trehalose and sucrose. The dehydrated trehalose matrix is homogeneous in terms of distribution of the residual water and spin-probe molecules. In contrast, dehydrated sucrose forms a heterogeneous matrix. It is comprised of sucrose polycrystalline clusters and several bulk water domains. The amorphous form was found only in 30% (volume) of the sucrose matrix. Re-hydration leads to a structural homogenization of the sucrose matrix, whilst in the trehalose matrix several domains develop differing in the local water/radical content and radical mobility. The molecular model of the matrices provides an explanation for the different protein-matrix dynamical coupling observed in dried ternary sucrose and trehalose matrices, and accounts for the superior efficacy of trehalose as a bioprotectant. Furthermore, for bacterial photosynthetic reaction centers it is shown that at low water content the protein-matrix coupling is modulated by the sugar/protein molar ratio in sucrose matrices only. This effect is suggested to be related to the preference for sucrose, rather than trehalose, as a bioprotective disaccharide in some anhydrobiotic organisms.

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“…Trehalose, a natural non‐reducing disaccharide present in diverse organisms, is widely used in the food and drug storage industry 36,37 . To the best of our knowledge, only a few studies have assessed the effect of trehalose on bone mass.…”

Section: Discussionmentioning

confidence: 99%

Trehalose reduces bone loss in experimental biliary cirrhosis rats via ERK phosphorylation regulation by enhancing autophagosome formation

Wang

et al. 2020

FASEB j.

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Bone loss is a severe complication of primary biliary cirrhosis (PBC). Trehalose was intermittently administered in bile duct‐ligated (BDL) male rats, a PBC‐related osteoporosis model, for 4 weeks to reduce osteoporosis. Femoral bones were assessed ex vivo by micro computed tomography (CT) and histomorphometry. The potential mechanisms related to the reduction of osteoporosis were explored by evaluating the effect of trehalose on osteoblast autophagy, osteogenesis, osteoclastogenesis, and ERK phosphorylation. The results demonstrated that trehalose reduced osteoporosis of BDL rats and decreased osteoblast‐mediated osteoclast differentiation by enhancing osteoblast autophagy to regulate osteoprotegerin (OPG) secretion. Hydroxychloroquine (HCQ) increased the expression of OPG and OPG/receptor activator genes for nuclear factor‐κB ligand (RANKL) ratio, and reduced osteoblast‐mediated osteoclastogenesis by inhibiting autophagy flux and inducing autophagosome formation. Furthermore, trehalose increased the phosphorylation of ERK1/2 in MC3T3‐E1 cells, and the ERK inhibitor PD98059 reversed the upregulation of OPG gene and reduction of trehalose‐induced osteoclastogeneis. The treatment with HCQ markedly increased the ERK phosphorylation. The correlation between autophagosome formation and ERK phosphorylation was confirmed in autophagy proteins (ATG) 4B or ATG5‐deficient cells. Thus, trehalose could decrease osteoblast‐mediated osteoclastogenesis and reduce PBC‐related bone loss by regulating ERK phosphorylation via autophagosome formation.

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Trehalose Preserves the Integrity of Lyophilized Phycoerythrin–AntiHuman CD8 Antibody Conjugates and Enhances their Thermal Stability in Flow Cytometric Assays

Cited by 12 publications

References 48 publications

The Magic of Disaccharide Glass Matrices for Protein Function as Decoded by High-Field EPR and FTIR Spectroscopy

The Magic of Disaccharide Glass Matrices for Protein Function as Decoded by High-Field EPR and FTIR Spectroscopy

Structural and dynamical characteristics of trehalose and sucrose matrices at different hydration levels as probed by FTIR and high-field EPR

Trehalose reduces bone loss in experimental biliary cirrhosis rats via ERK phosphorylation regulation by enhancing autophagosome formation

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