Protein cell free system. Medical search. Frequent questions

The NEBExpress Cell-free E.coli Protein Synthesis System can synthesize protein in just 2 to 4 hours. (neb.com)
Once the template DNA is purified, thaw the NEBExpress Cell-free E. coli Protein Synthesis System components on ice. (neb.com)
Recombinant expression of proteins of interest in Escherichia coli is an important tool in the determination of protein structure. (iucr.org)
An impediment to the successful production of large quantities of natively folded proteins in Escherichia coli is the tendency of many proteins to become insoluble when overexpressed. (iucr.org)
Protein aggregation remains a significant problem in E. coli expression systems. (iucr.org)
Common cell extracts in use today are made from E. coli (ECE), rabbit reticulocytes (RRL), wheat germ (WGE), insect cells (ICE) and Yeast Kluyveromyces (the D2P system). (wikipedia.org)
The biotechrabbit RTS 100 E. coli Disulfide Kit provides an optimized combination of lysate, reagents, chaperones, and buffer systems to produce maximum yields of active proteins containing multiple disulfidebonds in a CECF system. (biotechrabbit.com)
The RTS E. coli Disulfide Kit is designed for cell-free high-throughput expression of disulfide-bonded proteins from plasmid templates. (biotechrabbit.com)
Based on product type, the market has been segmented into E. coli cell-free protein expression system, rabbit reticulocytes cell-free protein expression system, wheat germ cell-free protein expression system, insect cells cell-free protein expression system, mammalian cell-free protein expression system, and consumables (labelling tags, vectors). (medgadget.com)
C ) GFVA-RG and GFVA-RS expressed using E. coli were purified using Ni beads and mixed with His-bls-FLAG-GST, which was synthesized using a wheat cell-free system and purified using glutathione beads. (elifesciences.org)
CFE reactions based on the use of Escherichia coli can quickly synthesize grams of protein per liter in batch reactions, are scalable from the nanoliter to 100-L scale, and can be freeze-dried for months of shelf stability. (scienceboard.net)
This work leveraged the E. coli -based CFE system to make one dose of vaccine an hour, costing around $5 per dose -- not an optimal cost for population-based vaccination programs. (scienceboard.net)
Specifically, by using lipid nanodiscs as cellular membrane mimics, we obtained yields of up to 440 µg/mL for the single-subunit OST enzyme, 'Protein glycosylation B' (PglB) from Campylobacter jejuni , as well as for three additional PglB homologs from Campylobacter coli, Campylobacter lari , and Desulfovibrio gigas . (biorxiv.org)
Presented here is a protocol for the preparation and buffer calibration of cell extracts from exonuclease V knockout strains of Escherichia coli BL21 Rosetta2 ( ΔrecBCD and ΔrecB ). (jove.com)
We present the preparation steps of cell lysate from Escherichia coli BL21 Rosetta2 ΔrecBCD strain by sonication lysis and buffer calibration for Mg-glutamate (Mg-glu) and K-glutamate (K-glu) specifically for linear DNA. (jove.com)
This method is able to achieve protein expression levels comparable to that from plasmid DNA in E. coli CFPS. (jove.com)
In this protocol, steps for the preparation of cell-free lysates from the E. coli BL21 Rosetta2 ΔrecBCD strain by sonication lysis are described in detail. (jove.com)
Motivated by a minimal model of cell-free metabolism, this work explored the effects of energy molecules, which power TX-TL, and fuel molecules, which regenerate energy by harnessing core metabolism, on an E. coli -based TX-TL system. (biorxiv.org)
E. coli, P. pastoris , Mammalian cells, Plant cells and in vitro by cell free synthesis. (lu.se)

The synthesized protein can be isolated by affinity purification, for example, using NEBExpress Ni-NTA magnetic beads or NEBExpress Ni Spin columns for further analysis. (neb.com)
The market has been segmented based on major applications such as enzyme engineering, protein labelling, protein-protein interaction, and protein purification. (medgadget.com)
A His -or Strep-tag II can be added to either terminus, greatly simplifying protein purification and detection after expression. (qiagen.com)
Increasing toolbox to approach the production and purification of difficult-to-express proteins to fulfill the demand for difficult-to-express proteins, which include different expression systems, promoters that have different strengths, cultivation media, and conditions. (futuremarketinsights.com)
For added convenience, at family of pT7CFE vectors are available with tandem affinity tags for facilitate protein detection and purification. (thermofisher.com)
For Objective 3, the initial steps of developing a cell free culture system will involve optimization of purification and cell free storage of E. ruminantium and validation of methods to measure protein incorporation that do not require radioactive materials. (usda.gov)

These data indicate that this eukaryotic cell-free expression system has a high success rate and that the addition of specific reagents can increase the yield of soluble protein. (iucr.org)
These results are compatible with a model in which proteins of the sarcomere are in kinetic equilibrium with homologous proteins in a soluble pool. (rupress.org)
However it has proved challenging to identify a heterologous expression platform that can produce a soluble protein-based vaccine in a manner compliant with current Good Manufacturing Practice (cGMP). (nature.com)
This work identifies Drosophila S2 cells as a clinically-relevant platform suited for the production of 'difficult-to-make' proteins from Plasmodium parasites, and identifies a PfRH5 sequence variant that can be used for clinical production of a non-glycosylated, soluble full-length protein vaccine immunogen. (nature.com)
For example, a protein with several membrane-spanning domains that may or may not enter into the heterologous host's membrane bilayers or a protein that cannot be produced in a soluble form. (futuremarketinsights.com)
Transforming growth factor-alpha (TGF-alpha) is synthesized and transported to the cell surface as a membrane-anchored precursor (proTGF-alpha) that is converted to the soluble form by proteolytic cleavage. (nih.gov)
In the cell-free reconstitution system, the membrane-bound Suc-Ala-Ala-Ala-MCA-hydrolyzing enzyme from CHO cells was also activated by phorbol ester in the presence of the soluble fraction, ATP, and vanadate. (nih.gov)

Other factors driving cell free protein expression market are a growing demand for simple and efficient protein production methods, contamination free approach and increasing focus on production of mammalian cell free lysate due to drug discovery. (medgadget.com)
Mammalian cell-free protein expression system segment is expected to grow with the fastest CAGR over the forecast period, owing to increasing usage of mammalian cell lysate in humanized proteomic and biologics study. (medgadget.com)
These issues stem largely from the fact that cell lysate contains an active and complex metabolism whose effect on TX-TL has remained largely uncharacterized. (biorxiv.org)

As a control, the expression levels of enzyme-fused protein and target proteins were detected using each protein-specific antibody (left panel). (elifesciences.org)
A particular challenge is the synthesis of oligosaccharyltransferases (OSTs), which catalyze the attachment of glycans to specific amino acid residues in target proteins. (biorxiv.org)
Furthermore, we demonstrate the ability of cell-free synthesized OSTs to glycosylate multiple target proteins with varying N -glycosylation acceptor sequons. (biorxiv.org)
Proteins that activated immunity are potential antigens for immunization and the multi-omics phylum-spanning prioritization database that was created is a valuable resource for identifying target proteins in a wide array of different parasitic nematodes. (frontiersin.org)

The antibodies induced by one protein variant were shown to be qualitatively similar to responses induced by other vaccine platforms. (nature.com)
Anti-merozoite vaccine studies have long relied on the standardized in vitro assay of growth inhibition activity (GIA) 12 , whereby purified IgG antibodies are tested against parasites cultured in human red blood cells (RBC) in the absence of other cell types. (nature.com)
Using antibodies to IDO, they didn't find any IDO expression until they treated the cells with interferon to mimic the conditions in the lymph nodes. (sciencedaily.com)
In this research project, we have investigated the C-terminal structures of small G proteins, purified the regulatory proteins for small G proteins, and made monoclonal antibodies against small G proteins and their regulatory proteins. (nii.ac.jp)
Among these regulatory proteins, we have cloned the cDNAs of smg p21 GDS, smg p25A GDI, and rho GDI, and made monoclonal antibodies against them. (nii.ac.jp)
Breastmilk has cells, hormones, and antibodies that help protect your baby from illnesses. (cdc.gov)

Screening the array with sera and ileal fluid samples from immunized pigs suggested cross-reactivity among homologous proteins and a general activation of immunity. (frontiersin.org)

The difficult-to-express proteins market is likely to register a CAGR of 11.33% during the forecast period and is anticipated to reach a market share of US$ 12.37 Billion in 2032, from US$ 3.8 Billion in 2021, due to the development of new tools and significantly boost the overall revenue. (futuremarketinsights.com)
In heterologous expression systems, many proteins are difficult to express, stumbling the demand for difficult-to-express proteins. (futuremarketinsights.com)
Furthermore, the target protein's inherent features provide difficulties for the expression host, impacting the sales of difficult-to-express proteins. (futuremarketinsights.com)
Many proteins require post-translational modifications that may or may not be present in each host, influencing the difficult-to-express proteins market. (futuremarketinsights.com)
In addition, the development of other microbial cell factories towards their use as effective protein production platforms is expected to spur the global sales of difficult-to-express proteins. (futuremarketinsights.com)
What Contributes to the Growth of the Difficult-To-Express Proteins Market? (futuremarketinsights.com)
The development of new tools for better difficult-to-express proteins market share will greatly increase the overall sales of difficult-to-express proteins. (futuremarketinsights.com)
Development of innovative cell-free protein synthesis platforms based on the industrial workhorse CHO cells that address the difficulties that continue to drive the demand for difficult-to-express proteins. (futuremarketinsights.com)
Prokaryotic expression systems will continue to play an essential role as a cost-effective, customizable, and simple-to-run source of recombinant protein as the number of authorized therapeutic proteins grows, encouraging the sales of difficult-to-express proteins. (futuremarketinsights.com)
What are the Restraining Factors in the Difficult-To-Express Proteins Market? (futuremarketinsights.com)
However, some obstacles that may limit the demand for difficult-to-express proteins include the high cost of systems and the difficulty of analyzing the target end-user segment, as systems for difficult-to-express proteins are confined to end users. (futuremarketinsights.com)
Which is the Key Region in the Difficult-To-Express Proteins Market? (futuremarketinsights.com)
Due to increased research and development, and collaboration between pharmaceutical firms and university research institutes for the development of novel tools and technologies, North America is likely to contribute significant difficult-to-express proteins market share. (futuremarketinsights.com)
The sales of difficult-to-express proteins will be boosted by increasing research in the field of protein expression and the adoption of new techniques for various diagnostic purposes. (futuremarketinsights.com)
The demand for difficult-to-express proteins in Asia Pacific is primarily driven by factors such as the growing presence of large biotechnology and biomedical businesses in the region. (futuremarketinsights.com)

Thus, eukaryotic in vitro systems, specifically the wheat germ cell-free system, hold significant promise. (iucr.org)
Wheat germ cell-free protein expression system is also anticipated to witness a CAGR of 5.8% over the forecast period. (medgadget.com)

Co-immunoprecipitation and mass spectrometric evaluation of proteins from rgCRFP-infected hen embryo fibroblasts utilizing business anti -RFP antibody led to the identification of a complete of 21 gC interacting host proteins. (eqalix.com)
The researchers used this process to develop a protein vaccine candidate modified with a sugar structure that could trigger the immune system, as well as a therapeutic antibody fragment with a sugar that can stabilize proteins as they circulate in the body. (northwestern.edu)
A cocktail of five recombinant proteins optimized for conserved GI nematode targets was used immunize pigs and test for active antibody responses in both the serum and intestinal ileal fluid of immunized pigs. (frontiersin.org)
An antibody-protein array of putative immunogenic proteins was developed from a combined bioinformatic, experimental, and literature-based prioritization of homologous parasite proteins. (frontiersin.org)
We have also found that smg p25A is mainly detected inneural tissues by use of its specific monoclonal antibody, and that the levels of smg p25A mRNA increase in PC12 cells after their differentiation into neuron-like cells. (nii.ac.jp)
Antibody -- Any of the protein molecules produced by specialized immune system cells (B cells) that can recognize and bind to a particular foreign antigen. (nih.gov)

Three possible PR/nanoparticle formation strategies were analyzed: (i) PR integration into supplied preassembled nanoparticles, (ii) coassembly of nanoparticles from supplied scaffold proteins and lipids upon PR expression, and (iii) coexpression of scaffold proteins together with PR in presence of supplied lipids. (moeller-lab.com)
Yes, ANXA4 has been shown to interact with several proteins and lipids. (creativebiomart.net)

CFPS has many advantages over the traditional in vivo synthesis of proteins. (wikipedia.org)

April 22, 2021 -- A new cell-free synthetic biology technique that increases the volume of membrane components in the manufacture of conjugate vaccines can significantly improve the efficiency of vaccine production, according to a new method published in Nature Communications on April 22. (scienceboard.net)
Beginning with the 2021-2022 season, all four flu viruses used in the cell culture-based vaccine are grown in cell cultures only, making the vaccine completely egg-free. (cdc.gov)

We also offer solutions for automation, site-directed mutagenesis, as well as your favorite restriction enzyme, ligase or competent cell products. (neb.com)
But because researchers have used mammalian cells to create the biosynthetic pathways (sets of enzyme catalysts) to build and study these sugar structures, the process has historically taken a long time and has required specialized laboratory equipment. (northwestern.edu)
The Jefferson scientists wanted to know if metastatic pancreatic cancer cells residing in the lymph nodes expressed IDO to avoid being found, and if so, could they target this enzyme with available drugs to prevent the cancer cells from hiding from the immune system. (sciencedaily.com)
The tumor cells were then able to make the enzyme. (sciencedaily.com)
Using the fluorogenic substrate succinyl-Ala-Ala-Ala-4-methylcoumaryl-7-amide (Suc-Ala-Ala-Ala-MCA), containing the sequence similar to the cleavage sites for proTGF-alpha processing, we identified a putative candidate proTGF-alpha-converting enzyme in the membrane fractions of Chinese hamster ovary (CHO) cells. (nih.gov)
The enzyme activity was about 5-6-fold increased by exposure of the cells with phorbol ester or with calcium ionophore. (nih.gov)
The enzyme was partially purified from membranes of phorbol ester-treated CHO cells. (nih.gov)
Inhibitor profile of the enzyme, including inhibition by serine protease inhibitors and no inhibition by elastatinal, coincides with that previously reported for proTGF-alpha cleavage activity on CHO cells. (nih.gov)
Requirement of vanadate, an inhibitor for protein phosphatases, for activation of the enzyme in this system suggests that phosphorylation-dephosphorylation probably plays a key role in the regulated cleavage of proTGF-alpha. (nih.gov)
Chymotrypsin -- An enzyme that catalyses the hydrolysis (the breakdown) of proteins into peptides or amino acids in the small intestine. (nih.gov)
Enzyme -- A protein molecule that catalyzes chemical reactions of other substances without itself being destroyed or altered by the reactions. (nih.gov)

After transfecting for 24 hr, cells were lysed by RIPA buffer including protease inhibitors, and biotinylated proteins were pulled down with streptavidin beads. (elifesciences.org)
2017). Determining the topology of peroxisomal proteins using protease protection assays . (up.pt)

Nanoparticles composed of amphiphilic scaffold proteins and small lipid bilayers are valuable tools for reconstitution and subsequent functional and structural characterization of membrane proteins. (moeller-lab.com)
Publications] Kaibuchi,K.: 'Molecular cloning and characterization of the stimulatory GDP/GTP exchange protein(GDS)for smg p21s,ras p21ーlike small GTPーbinding proteins. (nii.ac.jp)
2007). Proteomics characterization of mouse kidney peroxisomes by tandem mass spectrometry and protein correlation profiling . (up.pt)

Glycosylation - the attachment of sugars to proteins - plays a critical role in both cellular function and in the development of therapeutics, like vaccines. (northwestern.edu)
Cell-free gene expression (CFE) systems activate transcription and translation using crude cellular components instead of living, intact cells. (scienceboard.net)
These membrane vesicles are responsible for important cellular functions such as activating cost-effective energy metabolism from oxidative phosphorylation in bacterial CFE systems. (scienceboard.net)
ANXA4 can also interact with other annexin proteins or cytoskeletal proteins, potentially contributing to its diverse cellular functions. (creativebiomart.net)
The human cellular prion protein (PrP C ) is a glycosylphosphatidylinositol (GPI) anchored membrane glycoprotein with two N-glycosylation sites at residues 181 and 197. (mdpi.com)
Anti-cell death is a technique adopted by multi-cellular organisms as a defensive and developmental mechanism (Franklin-Tong and Gourlay, pg 390). (essaywriter.org)
Using a laboratory technique created at Johns Hopkins Medicine in 2010, the study researchers replicated the cellular environment in which specialized immune cells called antigen presenting cells (APCs) break down proteins derived from HIV and make them visible ("presented") to the immune system's frontline of defense, cells known as CD4+ T lymphocytes, or helper T cells. (technologynetworks.com)
[ 2 ] Oncocytic cells in the thyroid are often called Hürthle cells, and oncocytic change is defined as cellular enlargement characterized by an abundant eosinophilic granular cytoplasm as a result of accumulation of altered mitochondria. (medscape.com)

The advantages of using the 1-Step Human High-Yield IVT Kits over traditional in vivo expression systems include the ability to express toxic or insoluble proteins, easily perform protein labeling with modified amino-acids and reduce the time and cost of expressing human proteins in tissue culture cells. (thermofisher.com)

The dominant discovery in these experiments indicates that there is a heterologous expression of certain genes in plants that can regulate cell death. (essaywriter.org)

Cell-free protein synthesis, also known as in vitro protein synthesis or CFPS, is the production of protein using biological machinery in a cell-free system, that is, without the use of living cells. (wikipedia.org)
The in vitro protein synthesis environment is not constrained by a cell wall or homeostasis conditions necessary to maintain cell viability. (wikipedia.org)
Cell-free system -- A mixture of cytoplasmic and/or nuclear components from cells used for in vitro protein synthesis, transcription, DNA replication, or other purposes. (nih.gov)

The virus' hemagglutinin (HA) surface proteins are depicted in blue. (cdc.gov)

2017). The peroxisomal matrix protein translocon is a large cavity-forming protein assembly into which PEX5 protein enters to release its cargo . (up.pt)

Thus, endomembrane-modifying proteins within the endomembrane system will have an influence on the final grain quality/yield and recombinant protein production. (nature.com)

degradation is considered mediated by the interaction between the ubiquitin -proteasome system (UPS) and the autophagic pathway. (eqalix.com)
The protein-protein interaction segment is expected to register the highest CAGR of 6.0% over the forecast period due to increasing number of proteomic studies. (medgadget.com)
Does ANXA4 have any interaction partners or binding proteins? (creativebiomart.net)
Publications] Araki,S.: 'Role of the Cーterminal region of smg p25A in its interaction with membranes and the GDP/GTP exchange protein. (nii.ac.jp)
It is also possible that UL33 contributes to the transient interaction of terminase with the portal protein, UL6, during packaging. (gla.ac.uk)

Currently, her projects mainly focus on studying BRCA2 and Tel1 related protein-protein interactions involved in DNA damage response pathway, using Cryo-EM and other biophysics methods. (imperial.ac.uk)
The authors include information on the devices that support spatial engineering, RNA switches and explore the early applications of synthetic biology in protein synthesis, generation of pathway libraries, and immunotherapy. (wiley-vch.de)
2013). A cargo-centered perspective on the PEX5 receptor-mediated peroxisomal protein import pathway . (up.pt)
2009). Mapping the cargo protein membrane translocation step into the PEX5 cycling pathway . (up.pt)

Conduct serum protein electrophoresis, immunofixation analyses, and kappa and lambda free light chain (FLC) assays in serum, to determine the age-adjusted prevalence and monoclonal protein size distribution of MGUS by ethnic/racial group. (cdc.gov)

This system includes all of the components needed for protein synthesis: a highly active cell extract, an optimized T7 RNA polymerase, an RNase inhibitor and a protein synthesis reaction buffer containing energy source, nucleotides, and amino acids. (neb.com)
Combine the following in a 1.5 ml microcentrifuge tube: 12 µl of NEBExpress S30 Synthesis Extract, 25 µl of 2X Protein Synthesis Buffer, 1µl of T7 RNA Polymerase, 1 µl of RNase Inhibitor, 250 ng of DNA template, and nuclease free water to total reaction volume of 50 µl. (neb.com)
Furthermore, inhibition of endogenous PKA by the heat-stable PKA inhibitor is sufficient to stimulate MAPK activity in these extracts in the absence of protein synthesis and without dependence on a functional Ras protein. (ox.ac.uk)
An inhibitor might be able to activate T-cells to kill cancer cells, for example. (sciencedaily.com)
We have purified and characterized several GDP/GTP exchange proteins (GDP dissociation stimulator (GDS) and GDP dissociation inhibitor (GDI)) and GTPase activating Proteins (GAP) for small G proteins. (nii.ac.jp)

The Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has recently emerged as a leading candidate antigen against the blood-stage human malaria parasite. (nature.com)
Our simple method, called reductionist cell-free antigen processing, reproduces in a test tube the complex events that occur in the human immune system as a response to antigens, foreign invaders to the body such as viruses like HIV," says senior study author Scheherazade Sadegh-Nasseri, Ph.D. , professor of pathology at the Johns Hopkins University School of Medicine. (technologynetworks.com)
When APCs chew up proteins from an antigen and present the fragments, known as antigenic epitopes, on their surface, the epitopes become visible to helper T cells and initiate an immune response. (technologynetworks.com)
If the antigen is on the surface of a cell, this binding leads to cell aggregation and subsequent destruction. (nih.gov)
In 1939-1940, the Rhesus (Rh) blood group system was discovered, leading to the development of minor antigen compatibility testing. (medscape.com)
Then, the virus-containing fluid is collected from the cells and the virus antigen is purified. (cdc.gov)

A cell extract is obtained by lysing the cell of interest and centrifuging out the cell walls, DNA genome, and other debris. (wikipedia.org)
For a variety of organisms, close to 30% of the genome is used to encode membrane proteins," said co-author Neha Kamat, PhD, assistant professor of biomedical engineering at McCormick. (scienceboard.net)
The text reviews the synthesis of DNA and genome engineering and offers a discussion of the parts and devices that control protein expression and activity. (wiley-vch.de)
Of the remaining HSV-1 proteins necessary for genome encapsidation, neither UL17 nor UL32 interacted with UL33. (gla.ac.uk)
explosion further, consider that a fictitious small genome with 2002) More recently and more dramatically, the potential for 260 genes would host the same number of combinations as cell state conversions is exemplified by the reprogramming of the number of atoms in the visible universe! (lu.se)

The CECF technology prolongs the reaction time up to 24 hours and, therefore, enhances protein yield. (biotechrabbit.com)
This continuous-exchange cell-free (CECF) system enables protein expression in an overnight incubation of up to 750 µg/mL. (thermofisher.com)

Developing these medicines has generally required many years and a laboratory with highly specialized equipment to grow mammalian cells. (northwestern.edu)
These mammalian cells naturally produce glycosylated proteins, but are slow-growing and can be difficult to engineer, limiting the number and diversity of glycosylation structures that can be built and tested. (northwestern.edu)
First, CDC or one of its laboratory partners, uses influenza viruses that have been grown in cultured mammalian cells to make CVVs, which are then provided to a vaccine manufacturer. (cdc.gov)
Next, the vaccine manufacturer inoculates the CVVs into cultured mammalian cells (instead of into chicken eggs) and allows the CVVs to replicate (i.e., make copies) for a few days. (cdc.gov)

The system routinely produces approximately 25 μg of protein per 50 μl reaction, or 1/2 mg/ml. (neb.com)
Common components of a cell-free reaction include a cell extract, an energy source, a supply of amino acids, cofactors such as magnesium, and the DNA with the desired genes. (wikipedia.org)
An energy source is an important part of a cell-free reaction. (wikipedia.org)
Most notably, a cell-free reaction, including extract preparation, usually takes 1 -2 days, whereas in vivo protein expression may take 1-2 weeks. (wikipedia.org)
In contrast, once DNA is inserted into live cells, the reaction cannot be accessed until it is over and the cells are lysed. (wikipedia.org)
The water sets off a chemical reaction that activates the cell-free system, turning it into a catalyst for making usable medicine when and where it's needed. (scienceboard.net)
In this system, protein expression is performed in a proprietary dialysis device that allows a continuous supply of nucleotides, amino acids and energy generating substrates into the reaction while removing inhibitors of proteins synthesis. (thermofisher.com)
The kits are optimized with Accessory Proteins and Reaction Mixes that support protein synthesis using a DNA template. (thermofisher.com)
Inside the cell, enzymes create RNA and DNA by facilitating the reaction of ribose with adenosine. (nih.gov)
Talk to your healthcare provider if you had an allergic reaction to the antibiotics gentamicin or ciprofloxacin, or chicken or egg protein. (cdc.gov)

Watch this tutorial explaining the streamlined workflow for our new NEBExpress ® Cell-free Protein Synthesis System to learn how you can easily synthesize your protein in as little as 2 to 4 hours. (neb.com)
He is working on developing a workflow for prototyping proteins using cell-free systems and machine learning. (imperial.ac.uk)
Identification, classification, and analysis of subsets of cells is easily accomplished with a suite of purpose-built software tools and guided workflow. (sartorius.com)
Workflow indicates typical steps taken for use of the Incucyte ® Cell-by-Cell Analysis Software Module to segment individual cells and classify population subsets using area, eccentricity or fluorescence intensity of labeled cells. (sartorius.com)

Posttranslational incorporation of contractile proteins into myofibrils in a cell-free system. (rupress.org)
The incorporation of newly synthesized protein into myofibrils has been examined in a cell-free system. (rupress.org)
Thus, CFPS enables direct access and control of the translation environment which is advantageous for a number of applications including co-translational solubilisation of membrane proteins, optimisation of protein production, incorporation of non-natural amino acids, selective and site-specific labelling. (wikipedia.org)
A major application of CFPS is incorporation of unnatural amino acids into protein structures (see expanded genetic code). (wikipedia.org)

There is extensive literature on the variables leading to insoluble recombinant expression of proteins. (iucr.org)

[ 5 ] The 2022 WHO classification of endocrine tumors has replaced the term Hürthle cell carcinoma with oncocytic carcinoma. (medscape.com)

Haptoglobin is a colorless protein of the alpha-globulin fraction of human serum. (medscape.com)
A total of 250 microliter of serum was used for each study subject to perform the analyses to detect and quantify M protein and FLC (described below). (cdc.gov)
For those individuals where an abnormal protein band or equivocal pattern with conventional agarose-gel electrophoresis was found, an additional 250 microliter serum was used to perform immunofixation for validation purposes (see below). (cdc.gov)
The normal reference range of 0.26 to 1.65 for the free-kappa-lambda ratio in the FLC assay reflects a higher serum level of free-lambda light chains than would be expected given the usual kappa-lambda ratio of two intact immuno¬globulins. (cdc.gov)

However, it is unknown how these strategies affect functional folding, oligomeric assembly and membrane insertion efficiency of cell-free synthesized membrane proteins. (moeller-lab.com)
The Thermo Scientific 1-Step Human High-Yield In Vitro Translation (IVT) Kits enable the expression of functional proteins with 10 to 100 times greater yield per mL than other mammalian IVT systems. (thermofisher.com)
The 1-Step High-Yield IVT System uses modified HeLa cell extracts to take advantage of the robust human translation machinery and generate functional full-length proteins. (thermofisher.com)
Since robust localisation of IRIS-1 in the nucleus is observed only in some nuclei, it can suggest that IRIS-1 is localising in the nucleus depending on the differentiation state of the cells or in a subset of cells with different functional relevance. (lu.se)

Sugar structures allow these proteins to remain stable while enabling them to perform tasks, like attack a cancer cell or retrain the immune system. (northwestern.edu)
A protein that helps prevent a woman's body from rejecting a fetus may also play an important role in enabling pancreatic cancer cells to evade detection by the immune system, allowing them to spread in the body. (sciencedaily.com)
Researchers found that the metastatic pancreatic cancer cells in the lymph nodes produce enough of the protein, IDO, to wall-off the immune system's T-cells and recruit cells that suppress the immune response to the tumor. (sciencedaily.com)
According to Jonathan Brody, Ph.D., assistant professor of Surgery at Jefferson Medical College of Thomas Jefferson University, one way that metastatic cancer cells can survive in nearby lymph nodes is by avoiding the immune system. (sciencedaily.com)
Scientists know that IDO shuts off tryptophan production in T-cells, putting them in a resting state, and recruits a different type of immune cell called T-regulatory cells, which can inhibit the immune system. (sciencedaily.com)
If cells are escaping the primary tumor and going into another environment such as the lymph nodes, what are they doing to evade detection by the immune system? (sciencedaily.com)
These data point to the fact that IDO may play a role in helping cancer cells avoid the immune system. (sciencedaily.com)
The immune system appears to have a balance that can allow cancer cells to grow but also can detect and destroy them, Brody explains. (sciencedaily.com)
If we can identify which epitopes are 'immunodominant' - the ones that elicit the strongest immune system response to the virus - then we may have the essential ingredients for the long-sought recipe to make an effective HIV vaccine," explains Sadegh-Nasseri. (technologynetworks.com)
If you are Rh negative and your unborn baby is Rh positive, your body's immune system may attack your baby's blood cells. (medlineplus.gov)
Antigens are features of the influenza virus that are recognized by the immune system and that trigger a protective immune response. (cdc.gov)
Because the mucosal immune system is a a major site of extrinsic immune stimuation, and celiac disease is associated with a potent stimulation of the gut immune system, elaborating both T cell and B cell mediated immune responses, typically associated with an increase in expansion of B cells and plasma cells within the gut the assays also include serologic testing for celiac disease. (cdc.gov)
There are many immune cells in the skin tissue just underneath the top layer of your skin. (cdc.gov)

Consequently, we anti cipated that sperm mitophagy may very well be reconstituted in a cell-free system consisting of permeabilized mammalian spermatozoa co-incubated with porcine oocyte extracts. (eqalix.com)
Previously pp60v-src, cyclin A, p39mos, and maturation-promoting factor (composed of Cdc2 and cyclin B) have been shown to activate mitogen-activated protein kinase (MAPK) and MAPK kinase (MEK) in cell-free extracts of Xenopus oocytes. (ox.ac.uk)
In addition, preincubation of extracts with cAMP-dependent protein kinase (PKA) blocks stimulation of MAPK by cyclin, p21V12ras, PKC, or pp60v-src, by at least 50%, but stimulation by c-Mos is unaffected. (ox.ac.uk)
However, linear DNA templates are rapidly degraded by exonucleases that are naturally present in the cell extracts. (jove.com)
By using cell extracts from exonuclease-deficient knockout cells, linear DNA templates remain intact without requiring any end-modifications. (jove.com)

What is a prenatal cell-free DNA (cfDNA) screening? (medlineplus.gov)
Prenatal cell-free DNA (cfDNA) screening is a blood test for pregnant women. (medlineplus.gov)
Obstetrical, perinatal and genetic outcomes associated with non-reportable prenatal cell free DNA screening results. (cdc.gov)

Wilson R , Plinston C , Hunter N , Casalone C , Corona C , Tagliavini F , Chronic wasting disease and atypical forms of bovine spongiform encephalopathy and scrapie are not transmissible to mice expressing wild-type levels of human prion protein. (cdc.gov)
2023). Defining RNA oligonucleotides that reverse deleterious phase transitions of RNA-binding proteins with prion-like domains. (upenn.edu)
Nevertheless, there is a strong correlation between the neurotoxicity caused by prion proteins and the blockade of their normal proteolysis. (mdpi.com)

The development of cell free systems and biochemical assays will be an important step in further characterising these proteins. (gla.ac.uk)
Using the celiac assays we are evaluating if there is any correlation between seropositivity for celiac disease and markers for plasma protein abnormalities. (cdc.gov)

In combination with cell-free protein production systems, nanoparticles can be used to cotranslationally and translocon independently insert membrane proteins into tailored lipid environments. (moeller-lab.com)
Here we report the production of full-length PfRH5 protein using a cGMP-compliant platform called ExpreS 2 , based on a Drosophila melanogaster Schneider 2 (S2) stable cell line system. (nature.com)
One of the major bottleneck in the cell free protein expression market is the low protein production volumes in the process. (medgadget.com)
The rapidly increasing demand for simpler, rapid and efficient protein production methods is boosting the cell-free protein expression market in research and development field. (medgadget.com)
Here, we address this challenge by establishing a bacterial cell-free protein synthesis platform that enables rapid production of a variety of OSTs in their active conformations. (biorxiv.org)
Her work here is focussed on providing structural and mechanistic insight into the activity of diadenylate cyclase DacA, an essential membrane protein responsible for the production of c-di-AMP in Staphylococcus aureus . (imperial.ac.uk)
The production of complicated therapies requires the use of recombinant proteins, which is a significant sector in the pharmaceutical industry. (futuremarketinsights.com)
Furthermore, maximum total mRNA and protein production occur in different and opposite concentration regimes of Mg +2 and 3-PGA. (biorxiv.org)
Under translation inhibition, transcriptional output was uniform across Mg +2 and 3-PGA concentrations, but in a translation-only system, maximum protein production occurred in the previously found optimal regime of Mg +2 and 3-PGA, suggesting a TX-TL trade-off. (biorxiv.org)
When a 'poly-U'-RNA having only uracil bases was added to the system, radioactive measurements indicated the production of protein-like molecules made up entirely of one single amino acid - phenylalanine. (news-medical.net)
Lund Protein Production Platform (LP3) has recently published its standard operating procedures for recombinant protein expression using the Baculovirus Expression Vector System (BEVS) in insect cells. (lu.se)
The egg-based production process begins with CDC or another laboratory partner in the WHO Global Influenza Surveillance and Response System providing private sector manufacturers with candidate vaccine viruses (CVVs) grown in chicken eggs per current FDA regulatory requirements. (cdc.gov)
There also is a cell culture-based production process for flu vaccines that was approved by FDA in 2012. (cdc.gov)
Cell culture-based flu vaccine production does not require chicken eggs because the vaccine viruses used to make vaccine are grown in mammalian cell cultures (no animals are harmed by this process). (cdc.gov)

The study gives a comprehensive guideline for the rapid synthesis of membrane protein/nanoparticle samples by different processes and identifies key parameters to modulate sample yield and quality. (moeller-lab.com)
The 1-Step Human High-Yield IVT Kits are cell-free protein expression systems that provide all of the essential components required for transcription and translation. (thermofisher.com)
Consequently, endomembrane-modifying proteins will influence the final grain quality and yield. (nature.com)

The simple presentation of cell-free protein expression system makes it easier to integrate them into high throughput platforms for efficient biologics and proteomics studies. (medgadget.com)

Luke completed his D.Phil in Clinical Medicine at Oxford University (Merton College), but his research career has predominantly focused on investigating protein function using structural biology as a means to provide a mechanistic understanding in life and in disease. (imperial.ac.uk)

The association was rapid (t1/2 less than 15 min) and, for most of the proteins, relatively temperature insensitive. (rupress.org)
This strategy enables rapid generation of protein/nanoparticle complexes by avoiding detergent contact of nascent membrane proteins. (moeller-lab.com)

This kit uses specially designed primers to amplify coding DNA sequence and supplement it with regulatory elements required for optimal transcription and translation in cell-free expression systems. (qiagen.com)

Complementary with the binding of SQSTM1 and VCP to sperm mitochondria, two sperm-borne pro-mitophagy proteins, parkin co-regulated gene product (PACRG) and spermatogenesis related 18, underwent localization modifications after extract coincubation, which had been according to their degradation noticed inside fertilized porcine oocytes. (eqalix.com)
Cell-free protein expression systems allow protein screening without necessitating a gene-cloning step thus enabling an accelerated process/product development pipelines which makes up for attractive opportunity for market players. (medgadget.com)
Cell-free transcription-translation (TX-TL) systems have been used for diverse applications, from prototyping gene circuits to providing a platform for the development of synthetic life, but their performance is limited by issues such as batch-to-batch variability, poor predictability, and limited lifetime. (biorxiv.org)
The UL33 gene of herpes simplex virus type 1 (HSV-1) encodes a 130 amino acid (aa) protein that is essential for the cleavage of concatemeric viral DNA into monomeric genomes and their packaging into preformed capsids. (gla.ac.uk)
Similarly, GATA-1 has been shown to induce lineage switching expression values even if, for simplicity, we assume only ``on'' of committed cells in hematopoiesis, first in cell lines (Kulessa and ``off'' states for each gene. (lu.se)
In reality, gene somatic cells to a pluripotent cell state by a handful of transcrip- expression is graded, making the potential gene expression tion factors (Takahashi and Yamanaka, 2006). (lu.se)

Milan Mrksich , the Henry Wade Rogers Professor of Biomedical Engineering, Chemistry, and Cell and Molecular Biology at Northwestern's McCormick School of Engineering, is a co-author on the paper. (northwestern.edu)
Monisha was a Marie Curie Sklodowska Fellow within the ITN Translocation project and was enrolled in a PhD program at the Institute for Cell and Molecular Biosciences (ICaMB), Newcastle University, UK. (imperial.ac.uk)
The student will be broadly trained in research involving bacterial cell culture, animal infection studies, molecular biology, host response assessment, field studies involving tick collections, tick rearing and manipulations, and tick-host-pathogen interactions. (usda.gov)
Biochimica et Biophysica Acta - Molecular Cell Research, 1793 (11), 1669 - 1675. (up.pt)
Biochimica et Biophysica Acta - Molecular Cell Research, 1863 (1), 139 - 147. (up.pt)
Biochimica et Biophysica Acta - Molecular Cell Research, 1863 (5), 814 - 820. (up.pt)
Biochimica et Biophysica Acta - Molecular Cell Research, 1773 (7), 1141 - 1148. (up.pt)
Biochimica et Biophysica Acta - Molecular Cell Research, 1823 (10), 1958 - 1966. (up.pt)
However, little is known about the molecular mechanism underlying endomembrane system remodeling during barley grain development. (nature.com)

Amino acid -- The basic building block of proteins and polypeptides. (nih.gov)

The researchers therefore developed a new manufacturing platform -- called in vitro conjugate vaccine expression (iVAX) -- that uses cell-free synthetic biology, a process in which researchers remove a cell's outer wall (or membrane) and repurpose its internal machinery. (scienceboard.net)
Finally, the need for live animals or cell culture to grow E. ruminantium limits the ability to produce an inactivated vaccine. (usda.gov)
However, on August 31, 2016, FDA issued an approval for Seqirus, the sole FDA-approved cell culture-based flu vaccine manufacturer in the United States, to begin using cell culture-grown CVVs. (cdc.gov)
Currently, cell culture-based manufacturing is used to make inactivated flu vaccines (e.g., flu shots), not LAIV (nasal spray flu vaccine). (cdc.gov)
Cell culture-based technology also has the potential for a faster start-up of the flu vaccine manufacturing process. (cdc.gov)
While viruses used in previous seasons' cell culture-based vaccine have been grown in cell cultures, prior to the 2019-2020 season, some of the viruses provided to the manufacturer had been first propagated in chicken eggs. (cdc.gov)

Since there is no need to maintain cell viability, toxic proteins can be produced. (wikipedia.org)
Some desired proteins and labeled proteins are toxic to cells when synthesized. (wikipedia.org)
Hürthle cells are observed in both neoplastic and nonneoplastic conditions of the thyroid gland (eg, Hashimoto thyroiditis , nodular and toxic goiter ). (medscape.com)

However, lack of expression and insolubility remain significant challenges to the expression and crystallization of these proteins. (iucr.org)
Due to the open nature of the system, different expression conditions such as pH, redox potentials, temperatures, and chaperones can be screened. (wikipedia.org)
in vitro transcription/translation protein expression products are manufactured using quality chemicals and materials that meet our high standards. (biotechrabbit.com)
The RTS disulfide system is the only in vitro protein expression system that can express properly folded disulfide-bonded proteins that cannot be produced under reducing conditions. (biotechrabbit.com)
Dr. Brody, Charles Yeo, M.D., Samuel D. Gross Professor and chair of Surgery at Jefferson Medical College and their co-workers analyzed IDO expression in 14 lymph nodes to which pancreatic cancer cells had spread and compared them to the primary tumors that had not spread in the same patients. (sciencedaily.com)
In every case, they found greater expression of the IDO protein in the cancerous lymph nodes. (sciencedaily.com)
The group also examined pancreatic cancer cell lines in the laboratory for IDO expression. (sciencedaily.com)
The Cell-Free Protein Expression Market will witness a CAGR of 6%, reaching US$ 268.4 Million between 2024. (medgadget.com)
In terms of revenue, the global cell free protein expression market is projected to register a CAGR of 6.0% over the forecast period owing to various factors, on which PMR offers detailed insights and forecasts. (medgadget.com)
The primary factors fuelling demand for cell free protein expression market are increasing R&D outsourcing by pharmaceutical and biotechnological companies, declining R&D productivity and patent cliff sales drop leading to increasing research intensity in the pharmaceutical sector and increasing expenditure on biosimilar development. (medgadget.com)
See Through Sample Of Cell Free Protein Expression Market Report! (medgadget.com)
Starting With The New Decade On A Diligent Note In The Cell Free Protein Expression Market? (medgadget.com)
This limitation makes cell free protein expression unsuitable for large industrial applications and is generally preferred in small R&D processes. (medgadget.com)
However, efforts are being made to overcome this constraint and make it a preferred expression system in industrial settings. (medgadget.com)
How About Obtaining Insights About The Region To Enter Concerning The Cell Free Protein Expression Market? (medgadget.com)
Press The "Purchase Now" Button To Have Our Cell Free Protein Expression Market Report! (medgadget.com)
Numbers in parentheses denote the stated expression levels of PrP c into the animal species and cell lines used. (cdc.gov)
Real-time quantitative PCR (RT-qPCR) analysis and sfGFP (superfolder green fluorescent protein) analysis reveal that AmtR directly and negatively regulates the transcription and expression of crn A, cre T, csh A, and hyu B. A growth test shows that C. glutamicum ATCC 14067 can use creatine or creatinine as a sole nitrogen source. (frontiersin.org)
We recommend to use the EasyXpress Linear Template Kit Plus to generate PCR products optimized for use in protein expression with the EasyXpress Insect Kit II . (qiagen.com)
How is the expression of the ANXA4 protein regulated? (creativebiomart.net)
This is a fast, easy, and direct approach for expression in cell-free protein synthesis systems using linear DNA templates. (jove.com)
All these strategies cost extra time and resources and are yet to obtain near-plasmid levels of protein expression. (jove.com)
RMM has a financial stake in Tierra Biosciences, a private company that makes use of cell-free technologies such as those described in this article for protein expression and screening. (biorxiv.org)
1-Step Human IVT Kits are optimized for use with the pT7CFE1 expression vector that utilizes the T7 viral promoter and an EMCV Internal Ribosome Entry Site (IRES) to facilitate high levels of in vitro protein expression. (thermofisher.com)
In paper II, we showed that silencing of CD59 expression in rat β cells significantly suppressed insulin secretion. (lu.se)
Induction of glucotoxicity in primary, healthy human islets led to a significant decrease in IRIS-1 protein-level expression. (lu.se)
We found that IRIS-1 expressing cells displayed significantly higher expression levels of Urocortin 3 and Pdx 1 (markers of mature β cells, which loss marks the beginning of β cells dedifferentiation), suggesting that IRIS-1 may be required for maintaining β cells identity and function. (lu.se)
Recently, a book chapter with protocols for the Baculovirus Expression Vector System in insect cells was published by LP3. (lu.se)

article{60924ccc-1642-427f-ace4-07a32acfecf0, abstract = {{We present a Monte Carlo study of a model protein with 54 amino acids that folds directly to its native three-helix-bundle state without forming any well-defined intermediate state. (lu.se)

The Incucyte ® Cell-by-Cell Analysis Software Module enables a range of applications for mixed cultures of of adherent and non-adherent cells. (sartorius.com)
No. BA-04867) is an add on module to Cell-by-Cell Analysis module and enables identification of label-free cells with morphologies of interest. (sartorius.com)

Northwestern Engineering researchers have now developed a quick, cell-free system to build and study these pathways. (northwestern.edu)
In just a few months, Kightlinger used the system to construct 37 pathways, creating 23 unique sugar structures, 18 of which have never been synthesized on proteins. (northwestern.edu)
Future works will use other pathways developed in this paper to create glycosylated protein vaccines and therapeutics that can target certain areas within the body. (northwestern.edu)
The main aim of this paper is to review the pathways that lead to cell death in plants with a purpose of identifying the anti-cell death genes. (essaywriter.org)
By around 1960, the basic chemical pathways through which DNA instructs protein synthesis had been established. (news-medical.net)
In terminally differentiated cell fate is coupled to appropriate regulation of the alternative cells, transcriptional networks must be stable and irreversible, pathways. (lu.se)

Synthetic biology has many other uses and is a bright future in fields such as protein evolution, nanomachines, nucleic acid circuits, and synthesis of virus-like particles for vaccines and drug therapy. (wikipedia.org)
Glycosylation is important in the development of protein medicines, which include everything from anti-cancer drugs like Herceptin to flu and tetanus vaccines. (northwestern.edu)
The system could also be used to provide modular, on-demand biomanufacturing platforms that provide medicines or vaccines in resource-limited settings. (northwestern.edu)
These systems are used as platforms for biomanufacturing and synthetic biology, with applications for producing shelf-stable antibacterial medicines and vaccines. (scienceboard.net)
In the current study, the researchers found that increasing vesicle concentration could be useful in making components for protein therapeutics such as conjugate vaccines, which work by attaching a sugar unit -- which is unique to a pathogen -- to a carrier protein. (scienceboard.net)
The process of creating cell culture-based flu vaccines involves several steps. (cdc.gov)
For more information, see CDC's Cell Culture-Based Flu Vaccines webpage . (cdc.gov)

Other factors have since been linked with the decision by plant cells to activate cell death, including cell survival signals, stress signals, developmental cues, pathogen recognition and metabolic state. (essaywriter.org)

These membrane vesicles have been identified as components that enhance bacterial CFE systems but have mostly been underappreciated. (scienceboard.net)

Immunofluorescence studies of virally infected cells revealed that UL15 was necessary for the localisation of the remaining terminase components (UL28 and UL33) to nuclear sites of viral DNA replication, where packaging occurs. (gla.ac.uk)
It is therefore not yet possible to conclude whether the observed interactions of UL33 with these four proteins are essential for viral DNA packaging. (gla.ac.uk)

Out of those 21 proteins, the interplay of seven host proteins with DEV gC protein was validated utilizing membrane-bound split- ubiquitin yeast two-hybrid system and bimolecular fluorescence complementation (BiFC) analyses. (eqalix.com)

Through the execution of this program, mobile elements, together with aggregates, in addition to broken organelles and a few proteins are encapsulated in particular vesicles referred to as autophagosomes and subsequently are degraded after fusion of autophagosomes with lysosomes. (eqalix.com)
The team developed a method that enhances membrane-dependent functions of cell-derived vesicles in CFE systems. (scienceboard.net)
Membrane structures are present in all CFE systems, as vesicles formed by fragmentation and rearrangement of cell membranes during cell lysis and extract preparation. (scienceboard.net)

Hellmich W, Pelargus C, Leffhalm K, Ros A, Anselmetti D. Single Cell Manipulation, Analytics and Label-free Protein Detection in Microfluidic Devices for Systems NanoBiology. (uni-bielefeld.de)
Single Cell Manipulation, Analytics and Label-free Protein Detection in Microfluidic Devices for Systems NanoBiology" in Microfluidic Applications in Biology: From Technologies to Systems Biology , Lion, N., Roussier, J. S., and Girault, H. eds. (uni-bielefeld.de)

Bronchoalveolar lavage (BAL) was analysed for total and differential cell count, cell viability, and lipid and protein content. (cdc.gov)
No statistically significant differences in total cell counts, differential counts, viability, protein and lipid measurements, or lung histology were recognized among the 4 groups. (cdc.gov)

And 7 host proteins have been additional validated to work together with DEV gC utilizing MbYTH and BiFC analyses. (eqalix.com)
notably, integration of proteomics data with in situ subcellular microscopic analyses showed a high abundance of cytoskeleton proteins associated with acidified PBs at the early development stages. (nature.com)

Frequently in use are nanoparticles assembled with engineered derivatives of either the membrane scaffold protein (MSP) or the Saposin A (SapA) scaffold. (moeller-lab.com)

Anatomy3

Organisms1

Chemicals and Drugs2

Phenomena and Processes4

Information Science2

Coli19

Purification6

Soluble7

Lysate3

Target proteins4

Antibodies6

Homologous proteins1

Difficult-to-express proteins15

Wheat germ cell-free2

Antibody6

Lipids2

Synthesis of proteins1

20212

Enzyme11

Protease2

Characterization3

Cellular8

Human proteins1

Heterologous expression1

Vitro protein synthesis3

Surface proteins1

Peroxisomal matrix protein1

Recombinant protein production1

Interaction5

Pathway4

Monoclonal1

Inhibitor5

Antigen6

Genome5

CECF2

Mammalian cells4

Reaction10

Workflow4

Incorporation4

Insoluble1

20221

Serum4

Functional4

Immune13

Extracts5

Prenatal cell-free3

Prion3

Assays2

Production14

Yield3

High-throughput1

Structural1

Rapid2

Transcription and translation1

Gene6

Molecular9

Amino acid1

Vaccine6

Toxic3

Expression28

Abstract1

Enables2

Pathways6

Vaccines7

Activate1

Bacterial1

Viral2

Membrane-bound1

Vesicles3

Detection2

Viability2

Analyses2

Scaffold1