scholarly journals Back Cover: Mix to Validate: A Facile, Reversible PEGylation for Fast Screening of Potential Therapeutic Proteins In Vivo (Angew. Chem. Int. Ed. 27/2013)

2013 ◽  
Vol 52 (27) ◽  
pp. 7044-7044
Author(s):  
Tae Hyung Kim ◽  
Magdalena Swierczewska ◽  
Yumin Oh ◽  
AeRyon Kim ◽  
Dong Gyu Jo ◽  
...  
Keyword(s):  
Therapeutic Proteins ◽  
Back Cover ◽  
Fast Screening

scholarly journals Rücktitelbild: Mix to Validate: A Facile, Reversible PEGylation for Fast Screening of Potential Therapeutic Proteins In Vivo (Angew. Chem. 27/2013)

2013 ◽  
Vol 125 (27) ◽  
pp. 7182-7182
Author(s):  
Tae Hyung Kim ◽  
Magdalena Swierczewska ◽  
Yumin Oh ◽  
AeRyon Kim ◽  
Dong Gyu Jo ◽  
...  
Keyword(s):  
Therapeutic Proteins ◽  
Fast Screening

Mix to Validate: A Facile, Reversible PEGylation for Fast Screening of Potential Therapeutic Proteins In Vivo

2013 ◽  
Vol 125 (27) ◽  
pp. 7018-7022 ◽  
Author(s):  
Tae Hyung Kim ◽  
Magdalena Swierczewska ◽  
Yumin Oh ◽  
AeRyon Kim ◽  
Dong Gyu Jo ◽  
...  
Keyword(s):  
Therapeutic Proteins ◽  
Fast Screening

scholarly journals Mix to Validate: A Facile, Reversible PEGylation for Fast Screening of Potential Therapeutic Proteins In Vivo

2013 ◽  
Vol 52 (27) ◽  
pp. 6880-6884 ◽  
Author(s):  
Tae Hyung Kim ◽  
Magdalena Swierczewska ◽  
Yumin Oh ◽  
AeRyon Kim ◽  
Dong Gyu Jo ◽  
...  
Keyword(s):  
Therapeutic Proteins ◽  
Fast Screening

scholarly journals The Instability of Dimeric Fc-Fusions Expressed in Plants Can Be Solved by Monomeric Fc Technology

2021 ◽  
Vol 12 ◽  
Author(s):  
Pia Gattinger ◽  
Shiva Izadi ◽  
Clemens Grünwald-Gruber ◽  
Somanath Kallolimath ◽  
Alexandra Castilho
Keyword(s):  
Monoclonal Antibodies ◽  
Fusion Proteins ◽  
Degradation Products ◽  
Therapeutic Proteins ◽  
Cost Effective ◽  
Full Length ◽  
Peptide Sequence ◽  
Reporter Protein ◽  
The Stability

The potential therapeutic value of many proteins is ultimately limited by their rapid in vivo clearance. One strategy to limit clearance by metabolism and excretion, and improving the stability of therapeutic proteins, is their fusion to the immunoglobulin fragment crystallizable region (Fc). The Fc region plays multiple roles in (i) dimerization for the formation of “Y”-shaped structure of Ig, (ii) Fc-mediated effector functions, (iii) extension of serum half-life, and (iv) a cost-effective purification tag. Plants and in particular Nicotiana benthamiana have proven to be suitable expression platforms for several recombinant therapeutic proteins. Despite the enormous success of their use for the production of full-length monoclonal antibodies, the expression of Fc-fused therapeutic proteins in plants has shown limitations. Many Fc-fusion proteins expressed in plants show different degrees of instability resulting in high amounts of Fc-derived degradation products. To address this issue, we used erythropoietin (EPO) as a reporter protein and evaluated the efforts to enhance the expression of full-length EPO-Fc targeted to the apoplast of N. benthamiana. Our results show that the instability of the fusion protein is independent from the Fc origin or IgG subclass and from the peptide sequence used to link the two domains. We also show that a similar instability occurs upon the expression of individual heavy chains of monoclonal antibodies and ScFv-Fc that mimic the “Y”-shape of antibodies but lack the light chain. We propose that in this configuration, steric hindrance between the protein domains leads to physical instability. Indeed, mutations of critical residues located on the Fc dimerization interface allowed the expression of fully stable EPO monomeric Fc-fusion proteins. We discuss the limitations of Fc-fusion technology in N. benthamiana transient expression systems and suggest strategies to optimize the Fc-based scaffolds on their folding and aggregation resistance in order to improve the stability.

scholarly journals An Overview on Spray-Drying of Protein-Loaded Polymeric Nanoparticles for Dry Powder Inhalation

Pharmaceutics ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 1032
Author(s):  
Tânia Marante ◽  
Cláudia Viegas ◽  
Inês Duarte ◽  
Ana S. Macedo ◽  
Pedro Fonte
Keyword(s):  
Spray Drying ◽  
Polymeric Nanoparticles ◽  
Therapeutic Proteins ◽  
Pulmonary Delivery ◽  
Non Invasive ◽  
Rapid Absorption ◽  
Technological Advances ◽  
First Pass

The delivery of therapeutic proteins remains a challenge, despite recent technological advances. While the delivery of proteins to the lungs is the gold standard for topical and systemic therapy through the lungs, the issue still exists. While pulmonary delivery is highly attractive due to its non-invasive nature, large surface area, possibility of topical and systemic administration, and rapid absorption circumventing the first-pass effect, the absorption of therapeutic proteins is still ineffective, largely due to the immunological and physicochemical barriers of the lungs. Most studies using spray-drying for the nanoencapsulation of drugs focus on the delivery of conventional drugs, which are less susceptible to bioactivity loss, compared to proteins. Herein, the development of polymeric nanoparticles by spray-drying for the delivery of therapeutic proteins is reviewed with an emphasis on its advantages and challenges, and the techniques to evaluate their in vitro and in vivo performance. The protein stability within the carrier and the features of the carrier are properly addressed.

Back Cover: In Vivo Gold Complex Catalysis within Live Mice (Angew. Chem. Int. Ed. 13/2017)

2017 ◽  
Vol 56 (13) ◽  
pp. 3724-3724 ◽  
Author(s):  
Kazuki Tsubokura ◽  
Kenward K. H. Vong ◽  
Ambara R. Pradipta ◽  
Akihiro Ogura ◽  
Sayaka Urano ◽  
...  
Keyword(s):  
Gold Complex ◽  
Back Cover

Biochemical and Functional Characterization of PEGylated rVWF.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1021-1021 ◽  
Author(s):  
Peter L. Turecek ◽  
Friedrich Scheiflinger ◽  
Juergen Siekmann ◽  
Katalin Váradi ◽  
H. Peter Matthiessen ◽  
...  
Keyword(s):  
Binding Capacity ◽  
Functional Characterization ◽  
Therapeutic Proteins ◽  
Molecular Size ◽  
Covalent Modification ◽  
Prolonged Survival ◽  
Control Group ◽  
Base Line ◽  
Secondary Rise

Abstract Covalent modification of therapeutic proteins by polyethylene glycol derivatives is an established method for improving pharmacokinetic properties of therapeutic proteins. Highly purified rVWF expressed in CHO cells, was chemically modified via PEGylation of lysine residues at mild alkaline pH with PEG succinimidyl succinate (linear 5 kDa PEG). Increasing the amount of PEG used in the coupling procedure, the molecular size of VWF increased, as demonstrated in SDS-PAGE and by agarose gel electrophoreses indicating an increase in size of the bands resembling the VWF multimers. PEGylation of rVWF reduced platelet-aggregating and collagen-binding functions by 60% (VWF:RCo activity) and 40% (VWF:CB activity). While FVIII-binding capacity, measured by a FVIII binding ELISA, was reduced and reduction correlated with the amount of PEG bound to VWF, there was almost no effect on FVIII binding affinity which remained in the same order of magnitude as measured with non-PEGylated VWF. Similar results were obtained when rVWF was PEGylated via carbohydrate moieties after oxidation and subsequent derivatization with monomethoxy-PEG hydrazide. PEGylated rVWF was applied to VWF-deficient mice at a dose of 40 VWF:Ag U/kg and plasma levels were monitored for up to 24 hours. As a control, non-modified rVWF was applied to the animals. PEGylated VWF had substantially prolonged survival in the circulation compared with non-modified rVWF with an increase of the AUC by a factor of >10. VWD mice substituted with human VWF show a secondary rise in FVIII bringing them into the FVIII levels measured in C57Bl/6 control mice. This secondary rise was sustained after treatment with PEGylated rVWF where FVIII levels above the starting level were measurable even 48 hours after injection while in the control group base line FVIII levels were reached already after 24 hours. rVWF is the largest protein ever PEGylated and PEGylation results in prolonged survival in the circulation while maintaining FVIII stabilizing functions of the VWF molecule in vivo.

Engineering human cells for in vivo secretion of antibody and non-antibody therapeutic proteins

2011 ◽  
Vol 22 (6) ◽  
pp. 924-930 ◽  
Author(s):  
David Sánchez-Martín ◽  
Laura Sanz ◽  
Luis Álvarez-Vallina
Keyword(s):  
Therapeutic Proteins ◽  
Human Cells
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