Abstract:In 1989, Manning, Patel, and Borchardt wrote a review of protein stability (Manning et al., Pharm. Res. 6:903-918, 1989), which has been widely referenced ever since. At the time, recombinant protein therapy was still in its infancy. This review summarizes the advances that have been made since then regarding protein stabilization and formulation. In addition to a discussion of the current understanding of chemical and physical instability, sections are included on stabilization in aqueous solution and the dri… Show more
“…Manning entitled "Stability of Protein Pharmaceuticals". 167 The importance of this paper in the field is evident from the fact that it has been cited over 700 times as of August 2015.…”
“…Manning entitled "Stability of Protein Pharmaceuticals". 167 The importance of this paper in the field is evident from the fact that it has been cited over 700 times as of August 2015.…”
“…1,2 Currently, production processes for biologics are designed to remove degradation products, 1,2 and the biologics are formulated to increase stability. 1,3-6 Identification of antibodies that are stable throughout expression, purification, formulation, storage, distribution and in vivo is expected to reduce development and production costs, increase product shelf-life and improve potency in vivo if the chemical liability affects target binding or antibody pharmacokinectics. 1-6 Thus, it is increasingly common to include pharmaceutical developability as a criterion in the selection of development candidates.…”
Section: Introductionmentioning
confidence: 99%
“…The reaction affects protein charge and structure because all four products contain an additional negative charge compared to the parental asparagine, and, in the case of iso-Asp formation, 11 the polypeptide backbone is altered as well as the side chain.10.1080/19420862.2018.1504726-F0001Figure 1.Asparagine side chain deamidation products and intermediates.Cyclization of asparagine to succinimide involves the loss of the amine group and is considered irreversible under physiological conditions. 3 Hydrolysis of succinimide results in reversible generation of aspartic acid and iso-aspartic acid. Asparagine deamidation can also result in generation of the epimers D-Asp and D-iso-Asp.…”
Section: Introductionmentioning
confidence: 99%
“…3 Hydrolysis of succinimide results in reversible generation of aspartic acid and iso-aspartic acid. Asparagine deamidation can also result in generation of the epimers D-Asp and D-iso-Asp.…”
Developability assessment of therapeutic antibody candidates assists drug discovery by enabling early identification of undesirable instabilities. Rapid chemical stability screening of antibody variants can accelerate the identification of potential solutions. We describe here the development of a high-throughput assay to characterize asparagine deamidation. We applied the assay to identify a mutation that unexpectedly stabilizes a critical asparagine. Ninety antibody variants were incubated under thermal stress in order to induce deamidation and screened for both affinity and total binding capacity. Surprisingly, a mutation five residues downstream from the unstable asparagine greatly reduced deamidation. Detailed assessment by LC-MS analysis confirmed the predicted improvement. This work describes both a high-throughput method for antibody stability screening during the early stages of antibody discovery and highlights the value of broad searches of antibody sequence space.
“…[4] However, therapeutic proteins are susceptible to proteolysis and denaturation, limiting their efficacy in the body. [5,6] To solve the delivery problem, protein delivery systems based on for instance polymeric nanoparticles, [7] hydrogels, [8,9] or lipid-based nanoparticles [10,11] have been developed. Also mesoporous silica nanoparticles (MSNs) have been studied as carriers for a variety of biomolecules including anticancer drugs, oligonucleotides, and proteins.…”
Protein delivery into the cytosol of cells is a challenging topic in the field of nanomedicine, because cellular uptake and endosomal escape are typically inefficient, hampering clinical applications. In this contribution cuboidal mesoporous silica nanoparticles (MSNs) containing disk‐shaped cavities with a large pore diameter (10 nm) are studied as a protein delivery vehicle using cytochrome‐c (cytC) as a model membrane‐impermeable protein. To ensure colloidal stability, the MSNs are coated with a fusogenic lipid bilayer (LB) and cellular uptake is induced by a complementary pair of coiled‐coil (CC) lipopeptides. Coiled‐coil induced membrane fusion leads to the efficient cytosolic delivery of cytC and triggers apoptosis of cells. Delivery of these LB coated MSNs in the presence of various endocytosis inhibitors strongly suggests that membrane fusion is the dominant mechanism of cellular uptake. This method is potentially a universal way for the efficient delivery of any type of inorganic nanoparticle or protein into cells mediated by CC induced membrane fusion.
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