Characterization of protein aggregation: The case of a therapeutic immunoglobulin

2007 ◽  
Vol 1774 (1) ◽  
pp. 146-153 ◽  
Author(s):  
Barthélemy Demeule ◽  
M. Jayne Lawrence ◽  
Alex F. Drake ◽  
Robert Gurny ◽  
Tudor Arvinte
Keyword(s):  
Protein Aggregation

scholarly journals Aggregation-prone peptides modulate interferon gamma functionality in naturally occurring protein nanoparticles

2019 ◽  
Author(s):  
José Vicente Carratalá ◽  
Olivia Cano-Garrido ◽  
Julieta Sánchez ◽  
Cristina Membrado ◽  
Eudald Pérez ◽  
...  
Keyword(s):  
Biological Activity ◽  
Protein Aggregation ◽  
Interferon Gamma ◽  
Recombinant Proteins ◽  
Expression System ◽  
Protein Aggregates ◽  
Naturally Occurring ◽  
Protein Nanoparticles ◽  
Ifn Γ

AbstractEfficient protocols for the production of recombinant proteins are indispensable for the development of the biopharmaceutical sector. Approximately 400 recombinant protein-based biopharmaceuticals have been approved in recent decades, with steady growth projected in the coming years. During the expression of a heterologous gene, the protein quality control network is overcome by the disruption in protein homeostasis, leading to protein aggregation. This phenomenon has been described in all expression systems analyzed to date, including prokaryotic and eukaryotic host cells. These protein aggregates have long been considered inert protein clumps devoid of biological activity and their study has largely been neglected. However, in recent years, the classic view of protein aggregates has completely changed with the recognition that these aggregates are a valuable source of functional recombinant proteins. In this study, bovine interferon-gamma (rBoIFN-γ) was engineered to enhance the formation of protein aggregates by the addition of aggregation-prone peptides (APPs) in the generally recognized as safe (GRAS) bacterial Lactococcus lactis expression system. The L6K2, HALRU and CYOB peptides were selected to assess their intrinsic aggregation capability to nucleate protein aggregation. These APPs enhanced the tendency of the resulting protein to aggregate at the expense of the total protein yield. However, fine physicochemical characterization of the resulting intracellular protein nanoparticles (NPs), the protein released from these protein NPs, and the protein purified from the soluble cell fraction indicated that the compactability of protein conformations is directly related to the biological activity of variants of IFN-γ, which is used here as a model protein with therapeutic potential.ImportanceThe demand for recombinant proteins in the pharmaceutical industry is steadily increasing. Emerging novel protein formulations, including naturally occurring protein NPs, might be an alternative to soluble variants for fine analysis at the biophysical level. Such analyses are important to address safety about biological molecules.This study analyzes the effect of aggregation-prone peptides (APPs) on the improvement of the production of naturally occurring protein nanoparticles (NPs) of interferon gamma (IFN-γ) in the generally recognized as safe (GRAS) Lactococcus lactis expression system. In addition, the fine physico-chemical characterization of the resulting proteins, either obtained from the soluble or insoluble cell fractions, indicates that the selected engineered proteins embedded in the protein NPs show higher compactability than their soluble protein counterparts. Conformational compactability is directly related to the biological performance of the recombinant IFN-γ.

In-line characterization of a whey protein aggregation process: Aggregates size and rheological measurements

2013 ◽  
Vol 115 (1) ◽  
pp. 73-82 ◽  
Author(s):  
Fatou Toutie Ndoye ◽  
Nicolas Erabit ◽  
Denis Flick ◽  
Graciela Alvarez
Keyword(s):  
Protein Aggregation ◽  
Whey Protein ◽  
Aggregation Process ◽  
Rheological Measurements

scholarly journals E183K Mutation in Chalcone Synthase C2 Causes Protein Aggregation and Maize Colorless

2021 ◽  
Vol 12 ◽  
Author(s):  
Haixiao Dong ◽  
He Li ◽  
Yingjie Xue ◽  
Shengzhong Su ◽  
Shipeng Li ◽  
...  
Keyword(s):  
Protein Aggregation ◽  
Prokaryotic Expression ◽  
Chalcone Synthase ◽  
The Body ◽  
Ems Mutagenesis ◽  
Body Of Knowledge ◽  
Physiological Processes ◽  
Complete Protein ◽  
Whole Life Cycle

Flavonoids give plants their rich colors and play roles in a number of physiological processes. In this study, we identified a novel colorless maize mutant showing reduced pigmentation throughout the whole life cycle by EMS mutagenesis. E183K mutation in maize chalcone synthase C2 (ZmC2) was mapped using MutMap strategy as the causal for colorless, which was further validated by transformation in Arabidopsis. We evaluated transcriptomic and metabolic changes in maize first sheaths caused by the mutation. The downstream biosynthesis was blocked while very few genes changed their expression pattern. ZmC2-E183 site is highly conserved in chalcone synthase among Plantae kingdom and within species’ different varieties. Through prokaryotic expression, transient expression in maize leaf protoplasts and stable expression in Arabidopsis, we observed that E183K and other mutations on E183 could cause almost complete protein aggregation of chalcone synthase. Our findings will benefit the characterization of flavonoid biosynthesis and contribute to the body of knowledge on protein aggregation in plants.

Structural characterization of β-sheeted oligomers formed on the pathway of oxidative prion protein aggregation in vitro

2007 ◽  
Vol 157 (2) ◽  
pp. 308-320 ◽  
Author(s):  
Lars Redecke ◽  
Martin von Bergen ◽  
Joachim Clos ◽  
Peter V. Konarev ◽  
Dimitri I. Svergun ◽  
...  
Keyword(s):  
Protein Aggregation ◽  
Prion Protein ◽  
Structural Characterization

scholarly journals Characterization of Protein Aggregation via Intrinsic Fluorescence Lifetime

2009 ◽  
Vol 131 (46) ◽  
pp. 16608-16609 ◽  
Author(s):  
Kristian H. Schlick ◽  
Candace K. Lange ◽  
Gregory D. Gillispie ◽  
Mary J. Cloninger
Keyword(s):  
Protein Aggregation ◽  
Fluorescence Lifetime ◽  
Intrinsic Fluorescence

Characterization of Protein Aggregation Using Hydrogel-Encapsulated nIR Fluorescent Nanoparticle Sensors

ACS Sensors ◽  
2020 ◽  
Vol 5 (2) ◽  
pp. 327-337 ◽  
Author(s):  
Daniel P. Salem ◽  
Xun Gong ◽  
Heejin Lee ◽  
Alicia Zeng ◽  
Gang Xue ◽  
...  
Keyword(s):  
Protein Aggregation ◽  
Fluorescent Nanoparticle
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