Metabolomics analysis of soy hydrolysates for the identification of productivity markers of mammalian cells for manufacturing therapeutic proteins

2015 ◽  
Vol 31 (2) ◽  
pp. 522-531 ◽  
Author(s):  
Jason Richardson ◽  
Bhavana Shah ◽  
Pavel V. Bondarenko ◽  
Prince Bhebe ◽  
Zhongqi Zhang ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Therapeutic Proteins ◽  
Soy Hydrolysates ◽  
Metabolomics Analysis

scholarly journals RIPK1 regulates starvation resistance by modulating aspartate catabolism

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Xinyu Mei ◽  
Yuan Guo ◽  
Zhangdan Xie ◽  
Yedan Zhong ◽  
Xiaofen Wu ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Postnatal Period ◽  
Prenatal Period ◽  
Starvation Resistance ◽  
Atp Production ◽  
Metabolism Regulation ◽  
Autophagy Induction ◽  
Starvation Conditions ◽  
Metabolomics Analysis

AbstractRIPK1 is a crucial regulator of cell death and survival. Ripk1 deficiency promotes mouse survival in the prenatal period while inhibits survival in the early postnatal period without a clear mechanism. Metabolism regulation and autophagy are critical to neonatal survival from severe starvation at birth. However, the mechanism by which RIPK1 regulates starvation resistance and survival remains unclear. Here, we address this question by discovering the metabolic regulatory role of RIPK1. First, metabolomics analysis reveals that Ripk1 deficiency specifically increases aspartate levels in both mouse neonates and mammalian cells under starvation conditions. Increased aspartate in Ripk1−/− cells enhances the TCA  flux and ATP production. The energy imbalance causes defective autophagy induction by inhibiting the AMPK/ULK1 pathway. Transcriptional analyses demonstrate that Ripk1−/− deficiency downregulates gene expression in aspartate catabolism by inactivating SP1. To summarize, this study reveals that RIPK1 serves as a metabolic regulator responsible for starvation resistance.

scholarly journals Integrated use of LC/MS/MS and LC/Q-TOF/MS targeted metabolomics with automated label-free microscopy for quantification of purine metabolites in cultured mammalian cells

2018 ◽  
Author(s):  
S. Eric Nybo ◽  
Jennifer T. Lamberts
Keyword(s):  
Biological Samples ◽  
Mammalian Cells ◽  
Fluorescent Dyes ◽  
Neuroblastoma Cells ◽  
Digital Microscopy ◽  
Label Free ◽  
Targeted Metabolomics ◽  
Established Method ◽  
Purine Metabolites ◽  
Metabolomics Analysis

AbstractPurine metabolites have been implicated as clinically relevant biomarkers of worsening or improving Parkinson’s disease (PD) progression. However, the identification of purine molecules as biomarkers in PD has largely been determined using non-targeted metabolomics analysis. The primary goal of this study was to develop an economical targeted metabolomics approach for the routine detection of purine molecules in biological samples. Specifically, this project utilized LC/MS/MS and LC/QTOF/MS to accurately quantify levels of six purine molecules in samples from cultured N2a murine neuroblastoma cells. The targeted metabolomics workflow was integrated with automated label-free digital microscopy, which enabled normalization of purine concentration per unit cell in the absence of fluorescent dyes. The established method offered significantly enhanced selectivity compared to previously published procedures. In addition, this study demonstrates that a simple, quantitative targeted metabolomics approach can be developed to identify and quantify purine metabolites in biological samples. We envision that this method could be broadly applicable to quantification of purine metabolites from other complex biological samples, such as cerebrospinal fluid or blood.

scholarly journals Expression Systems and Species Used for Transgenic Animal Bioreactors

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Yanli Wang ◽  
Sihai Zhao ◽  
Liang Bai ◽  
Jianglin Fan ◽  
Enqi Liu
Keyword(s):  
Recombinant Proteins ◽  
Mammalian Cells ◽  
Mammalian Species ◽  
Therapeutic Proteins ◽  
Transgenic Animals ◽  
Mammary Glands ◽  
Transgenic Animal ◽  
High Fertility ◽  
Expression Systems ◽  
Specific Expression

Transgenic animal bioreactors can produce therapeutic proteins with high value for pharmaceutical use. In this paper, we compared different systems capable of producing therapeutic proteins (bacteria, mammalian cells, transgenic plants, and transgenic animals) and found that transgenic animals were potentially ideal bioreactors for the synthesis of pharmaceutical protein complexes. Compared with other transgenic animal expression systems (egg white, blood, urine, seminal plasma, and silkworm cocoon), the mammary glands of transgenic animals have enormous potential. Compared with other mammalian species (pig, goat, sheep, and cow) that are currently being studied as bioreactors, rabbits offer many advantages: high fertility, easy generation of transgenic founders and offspring, insensitivity to prion diseases, relatively high milk production, and no transmission of severe diseases to humans. Noticeably, for a small- or medium-sized facility, the rabbit system is ideal to produce up to 50 kg of protein per year, considering both economical and hygienic aspects; rabbits are attractive candidates for the mammary-gland-specific expression of recombinant proteins. We also reviewed recombinant proteins that have been produced by targeted expression in the mammary glands of rabbits and discussed the limitations of transgenic animal bioreactors.

scholarly journals Engineering Brain Parasites for Intracellular Delivery of Therapeutic Proteins

2018 ◽  
Author(s):  
Shahar Bracha ◽  
Karoliina Hassi ◽  
Paul D. Ross ◽  
Stuart Cobb ◽  
Lilach Sheiner ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Mammalian Cells ◽  
Neurological Diseases ◽  
Therapeutic Proteins ◽  
Intracellular Delivery ◽  
Protein Therapy ◽  
Secretion Systems ◽  
Intracellular Targeting ◽  
The Central Nervous System ◽  
Syndrome Protein

Protein therapy has the potential to alleviate many neurological diseases; however, delivery mechanisms for the central nervous system (CNS) are limited, and intracellular delivery poses additional hurdles. To address these challenges, we harnessed the protist parasite Toxoplasma gondii, which can migrate into the CNS and secrete proteins into cells. Using a fusion protein approach, we engineered T. gondii to secrete therapeutic proteins for human neurological disorders. We tested two secretion systems, generated fusion proteins that localized to the secretory organelles of T. gondii and assessed their intracellular targeting in various mammalian cells including neurons. We show that T. gondii expressing GRA16 fused to the Rett syndrome protein MeCP2 deliver a fusion protein that mimics the endogenous MeCP2, binding heterochromatic DNA in neurons. This demonstrates the potential of T. gondii as a therapeutic protein vector, which could provide either transient or chronic, in situ synthesis and delivery of intracellular proteins to the CNS.

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