Tag Archives: DNA

Inside the CSL, where Australia’s Oxford-AstraZeneca vaccine is made | Instant News

Behind a building in Melbourne’s northern suburbs is a long, narrow room roughly the size of a tennis court.

Technicians and operators – fully dressed in protective clothing, gloves, goggles, hairpins and masks – are busy walking around, handling a variety of shiny stainless steel tools that look like kilometers of plastic tubes.

There are people here all the time too – it’s a 24/7 operation.

That’s because this room, part of CSL’s Broadmeadows factory, is where 50 million doses of the Oxford-AstraZeneca COVID-19 vaccine will be planted.

Earlier this week, that vaccine approved by the Gene Technology Regulatory Office. Currently being reviewed in the Therapeutics Good Administration.

And as millions of Australians roll up their sleeves for the vaccine, the injection itself may only take a few seconds, but it will take three months to manufacture.

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The AstraZeneca vaccine is known as a biological vaccine. This requires the help of living organisms to be produced.

Many medicines are produced this way, including vaccines. The HPV vaccine, for example, which protects against cervical cancer, is one of them.

AstraZeneca’s COVID-19 vaccine relies on a type of cell called HEK 293.These cells were originally extracted from the kidneys of human embryos – hence HEK – in the 1970s. They grow well in the laboratory and are a component commonly used in medicine.

HEK cells don’t actually end up in vaccines. Instead, they developed an important part of the vaccine – the adenovirus, which carries the DNA spike protein blueprint.

So the first step in making the AstraZeneca vaccine is growing an army of HEK cells.

Just as you might get a sourdough starter from a friend, AstraZeneca, in November, supplied the CSL with tiny frozen tubes, each containing only milliliters of HEK cells.

CSL’s job is to take these bits and pieces and multiply them to fill the tiny water tank.

The CSL COVID-19 vaccine production facility has been working day and night to produce the AstraZeneca vaccine.(Science ABC: Belinda Smith)

And it’s not a simple matter of putting cells in a big tub, adding nutritional broth and other substances, and letting them grow.

HEK’s cells prefer company. They reproduce faster if they are busy with other people.

So the cells are started in a 10ml glass bottle. After they fill them, they are transferred to a slightly bigger one, maybe 50ml, and so on. (It’s a bit like a hermit crab growing bigger than its shell and finding a bigger one to transport.)

Finally, the cells are transferred into a plastic bag, like a larger version of the blood donation bag, and gently rocked on a mobile platform. The rotating motion helps them grow.

And finally, the now liter HEK cells are poured into the bioreactor: a 2,000 liter stainless steel barrel over 3 meters high.

After a few days, HEK’s cells, which live happily and reproduce in the bioreactor, are ready for the next step.

People in white lab coats and hairpins in rooms with stainless steel lab equipment
CSL has two bioreactors, each capable of accommodating 2,000 liters, at the COVID-19 vaccine production facility. It’s just using one for now.(Provided: CSL)

Introducing the adenovirus

At this point, about three weeks later, the adenovirus enters the process of making a vaccine – literally.

Also supplied in small tubes by AstraZeneca last November, adenovirus was added to awaiting bioreactor and HEK cells.

And while adenovirus is engineered so that it cannot replicate in human cells – it is missing the important genes that make it possible – HEK cells have been altered to encourage adenovirus to infect and replicate within them.

Two people in protective gear and laboratory equipment in stainless steel with the door open
The tools of these boxes are filters. Each batch of adenovirus vaccine is screened nine times.(Science ABC: Belinda Smith)

Over the next six days, adenovirus infected HEK cells, multiplied, and continued to infect more HEK cells.

HEK infected cells are sure to die. But after them, the number of adenoviruses increased dramatically.

Next, it’s time to filter and purify the adenovirus from the pale pink bioreactor broth.

This is done using a technique called capture chromatography.

The liquid drips through a membrane designed in such a way that the adenovirus sticks to it, but everything else flows right away.

The different fluid is then sent across the membrane, changing its charge so that it releases its grip on the adenovirus, which then collects.

The bulk concentrated adenovirus vaccine is frozen at -65 degrees Celsius in a 20 liter cryovault plastic container.

And their job is now done, HEK’s remaining cells are destroyed.

A man in a protective suit and hair clip in a cold room, holds the door open
Cryovault CSL keeps the vaccine concentrated at very low temperatures until it is time to dilute and mix it into the final product.(Provided: CSL)

Fill and finish (and test)

The first part – growing, harvesting and freezing – lasts for about six weeks on CSL’s Broadmeadows website.

The next six weeks involve diluting the vaccine to the correct concentration, packaging it up, tons of testing and preparing it for shipment to the clinic for use. This is called “fill and finish” and will take place at the Parkville facility, 14 km south of Broadmeadows, starting next week.

And while it sounds straightforward, it’s easier said than done.

Adenovirus vaccine can only last a long time if it is not refrigerated. So from the moment the block of vaccine ice is removed from deep freeze, the clock is ticking.

First, the cryovault blocks are thawed – again, on a rocking platform – at room temperature. This takes more than a day.

The liquid concentrate is then mixed with a buffer solution to produce about 200 liters, tested for concentration (which takes several hours) and then adjusted to levels set by AstraZeneca.

Why not inject adenovirus directly? Buffer solutions do a number of things, such as stabilizing the vaccine and maintaining its pH.

After the vaccines are mixed, they pass through two pharmaceutical filters to ensure that they are completely free of bacteria or other viruses.

The sterile liquid is eventually taken to the filling section, where 6.5ml – that is, 10 doses – are placed in a sterile glass bottle and covered with an aluminum liner and rubber stopper.

Glass bottle filled with clear liquid
Each bottle contains 10 doses of AstraZeneca vaccine.(Provided: CSL)

(The air above the filling machine is classified as “Grade A” air – the cleanest pharmaceutical grade air as defined by global regulations.)

It takes about 12 hours to put 200 liters into the bottle, and each batch ends up providing about 300,000 doses.

But that’s not the end of the process – not too far.

Then there are nearly two weeks of inspections and at least 14 tests – not including those conducted by the Therapeutic Goods Administration at the Canberra lab – and a final documentation check by AstraZeneca.

AstraZeneca, at last, gave the final green light to release a vaccine.

And then, after 12 weeks, it’s ready to go.

Safety and specifications

The entire vaccine process is carried out according to the strict specifications set by AstraZeneca.

Batches are tested at all stages, from raw materials, such as the chemicals used to grow HEK cells and the pouches they live in, and to contamination and purity.

Even quality control testers, who inspect each bottle after it is filled, undergo rigorous training beforehand. They were given a row of bottles to examine. Some have hidden defects.

They must find all defects three times to qualify.

And not all bottles are expected to make it through the mail. Between 2 and 5 percent may be rejected, based on the previous vaccine manufacturing process.

Glass bottle filled with clear liquid
Each bottle of AstraZeneca will be checked for defects. All who fail this test will be destroyed.(Provided: CSL)

Even though the production line is up and running in a few months – a process that usually takes the best time of a year and a half – these are still early days for CSL. The first few batches did not produce as many adenoviruses as expected.

But that’s the very nature of using living organisms, such as HEK cells, to make things. You just don’t know how they’ll act every time.

However, the production line at Broadmeadows continues. Multiple batches at various stages in the pipeline keep the product growing.

And once the first batch of vaccine is released, the company hopes to get more than a million doses out each week – with the potential to double that down the road.


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Why the US Needs to Improve Covid-19 Genome Sequencing | Instant News

The UK and South Africa discovered a new SARS-CoV-2 variant in their domestic Covid-19 cases. The variant was discovered using a genome sequencing technique that analyzes the structure of the virus and distinguishes mutations. This genome sequencing technique was regularly used around the world at the start of pandemics when we knew little about viruses, but it has been lost. The US and other countries will have to follow in the footsteps of Britain and South Africa when it comes to revamped genome sequencing regimes, as the next variant may be hiding in our backyard.

Genome sequencing essentially determines the chemical “base” sequence of the DNA molecule. Scientists use this sequence to identify genes, regulatory instructions, or in the case of Covid-19, mutations to the virus. Sequencing efforts at the start of the pandemic helped scientists determine the structure of the virus, as well as its initial mutations help a virus contagious enough to cause a massive pandemic.

Recently, genome sequencing was key to identifying the more transmissible variants found in the UK. The Covid-19 Genomics Consortium has traced the genetic history of Covid-19 for almost a year, 150,000 virus samples. While most viral variants have one or two minor mutations from each other, the British variants have 23 separate mutations. This discovery caused concern and further investigation by the Consortium, which determined that mutations caused the transmission process to accelerate. The British variant is thought to have become a massive fuel current cases in the UK in recent weeks.

South Africa’s new SARS-CoV-2 variant is was found with the same technique. The new strain was discovered at the end of November and announced a month later after further research and analysis. As found in the UK, this new strain was determined to be highly contagious compared to the strains we have handled for most of the pandemic. South Africa, which has weathered the Covid-19 storm relatively well, is now in the middle of a spike in a case like in England.

In the United States, our sorting efforts have dwindled over time. At the start of the pandemic, the global community was trying to find out what a virus was, and at that time many samples were genome sequenced in that effort. Currently, only 0.3% of the sample has been sorted in the United States, that is rating 43rd according to the GISAID Initiative, a global genome sequencing database project.

Sequencing can help fight Covid-19 and its emerging variants. British and South African variants have been detected in dozens case in the US. This transmission led me to believe that cases involving this variant were widespread, but the lack of genome sequencing allowed the variants to avoid surveillance. In response to this new variant, CDC announced a multiplication our sorting efforts.

The robust sequencing regime may find more than just additional cases of the British and South African variants. Viruses can mutate each time they infect a new host, and with tens of millions of cases recorded worldwide, there is likely a hidden variant waiting to be discovered. In the United States alone, several estimate indicates that 15-20% of Americans have contracted Covid-19, which would make a new variant originating from US cases very likely. If a domestically grown variant is out there, it might help with the recent spike in Covid-19 cases observed during the holiday season.

The prospect of a more contagious virus spreading in the US is scary, but it is possible that with enough mutations, a strain might be able to evade current vaccines. Therefore, efforts to find these strains must be strengthened. If a section of the population has vaccine-resistant strains of Covid-19, national public health agencies must identify them and continue vaccine research from there. The hope is that this mutation hasn’t happened yet, but we don’t know for sure.

Vaccines may include new strains in the UK and South Africa, vaccine distribution will continue as planned in the coming months, and life will return to normal in the second half of 2021. The opposite is also possible. Genome sequencing in the US and around the world must be supported, and then new strains must be identified and isolated. If not, we may see a very long year.

Full coverage and live updates about Coronavirus


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Dr Maha’s case: Three defendants refuse to provide DNA samples, police notifies court – Crime | Instant News

Published in 05 January 2021 5:00 p.m.

Dr Maha’s case: The three defendants refused to provide DNA samples, police notified the court

KARACHI (Dunya News) – Suicide investigative officer Dr Maha has informed the Karachi District Court that the three defendants in the case refused to provide DNA samples.

Dr Maha’s suicide case is tried at the Karachi City Court. The investigating officer files a report in court. The court was informed that the three suspects in the case were reluctant to provide DNA samples.

The court ordered investigating officers to submit a final challenge in five days.


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Breakthrough therapy is proven to shorten the time of human aging – BGR | Instant News

  • Scientists have developed a new therapy that they say can actually reverse aging at the genetic level.
  • By letting participants breathe pure oxygen in a hyperbaric chamber for five months a week for three months, the team can effectively reverse the age of chromosomal telomeres.
  • Obviously, other research is needed in this area, but so far, the results are promising.

Life as we know it follows an inevitable path that we call aging. Due to the nature of our DNA, humans have a limited lifespan (in terms of animal life on earth, it is actually all DNA). But by determining the way our DNA changes with age, and reversing the damage it causes over time, Israeli researchers seem to have taken a small step towards establishing a science-based fountain of youth step.

Such as telegraph According to reports, the team of scientists at Tel Aviv University focused on a part of chromosomes called telomeres. The telomeres at the ends of our chromosomes are longer when we are born, and become shorter over time. They are the protective features of our genes. As we age, they shorten, DNA damage will occur, and as a result our bodies begin to collapse.

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Therefore, researchers have proposed a novel treatment method to solve the problem of aging. They want to see if oxygen can reverse and restore DNA vitality, effectively shortening time by extending telomeres and enhancing their defense capabilities.

In the three-month trial, volunteers age 64 and older sat in a hyperbaric chamber and were asked to wear a mask to breathe. The mask provided 100% oxygen, and each participant sat indoors for 90 minutes five days a week. After the trial period ended, the researchers checked the participants’ DNA and found that the individual’s telomeres were “young” again. Scientists say that telomeres look like the participants in their 20s, which is indeed a remarkable achievement.

In addition, oxygen therapy can be used in combination with other types of anti-aging therapies to further increase the life span of animals including humans. There has not been any detailed research on the end results of using multiple therapies. Although the DNA of study participants does show specific benefits on a microscopic level, it is still unclear how these changes translate in terms of lifespan. Will these participants live longer? If so, how long will it take? Are there potential drawbacks to this therapy?

These and other questions need to be answered before anyone can claim that they have found a way to extend human life beyond what can be achieved by a healthy lifestyle alone.

Mike Wehner has covered technology and video games over the past decade, covering major news and trends in VR, wearables, smartphones and future technologies. Recently, Mike served as the technical editor of The Daily Dot, and has gained attention in USA Today, Time.com, and countless other online and print shops. His love of reporting is second only to his addiction to games.


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Simultaneous mutation of two nonessential genes can lead to the death of cancer cells | Instant News

Ludwig cancer research, has revealed a new instance in which simultaneous mutation of two nonessential genes;none of which are vital for the survival of cells can cause cancer cell death.

Headed by member of the Ludwig San Diego Richard Kolodner and published in the current The proceedings of the National Academy of Sciences, the study also shows that it is a deadly combination, or “synthetic lethality” can be reproduced in a drug-like molecule that can be used to treat cancer.

The development and FDA approval of a new generation of drugs called PARP inhibitors, for the treatment of malignant tumors with defects in tumor suppressor genes BRCA1 and BRCA2 that cause breast cancer, ovarian and many other cancers, have generated significant interest in using synthetic lethal interactions to develop cancer treatments.

Scientists, including group Kolodner, are on the hunt for other synthetic lethal interactions in cancers. “PARP inhibitors are a major step forward, but they are not perfect. Patients can become resistant to them, so there’s always a need for new and better treatments.”

Building from research done on yeast cells, Kolodner and his colleagues found that disabling or removing FEN1 gene of mammals, which is essential for DNA replication and repair, is fatal to cancer cells, mutated forms of the genes BRCA1 and 2.

We have provided information that should make people think FEN1 as a potential interesting therapeutic target and showed how yeast can be used to predict a number of synthetic lethal interactions, which can then be tested in a bona FIDE cancer cell lines with genetic instruments”.

Richard Kolodner, Professor, Professor, Department of cellular and molecular medicine, University of California, San Diego

In previous work with yeast Saccharomyces as a model to identify and study genes that maintain the integrity of the genome, Kolodner and his colleagues found that the RAD27 gene, and of synthetic lethal interactions with the 59 other nonessential genes of yeast.

Two such genes, it should be noted RAD51 and RAD52 play a role in recombination of DNA.

FEN1 is a close analogue or homologue, RAD27 in mammals. Based on their studies of yeast, Kolodner and his colleagues predicted that FEN1 synthetic lethal interactions with BRCA1 and BRCA2, which function in the same biochemical reactions in mammals, as RAD51 and RAD52 to do in yeast.

To test this hypothesis, they synthesized four FEN1-blocking molecules and used the best of them, S8, to suppress the activity of FEN1 in tumor cell lines with or without BRCA mutations. C8 proved to be an effective killer of BRCA-mutant cells.

Then they demonstrated that genetic disorders FEN1 expression had the same effect that S8 did for the breast cancer gene-mutant cells, confirming that the S8 worked, causing synthetic lethality.

Finally, the researchers instilled in C8-C8 sensitive and-resistant tumors in mice and showed that C8 significantly inhibited the growth of C8-sensitive tumors, but not in C8-resistant tumors.

Interestingly, not all cancer cell lines and tumors that responded to treatment C8 was deficient BRCA, K, indicating that FEN1 and synthetic lethal interactions with other genes as well.

These results reveal FEN1 as a novel target for drugs for the treatment of various malignant tumors by induction of synthetic lethality.

They also demonstrate that yeast-based screens provide a powerful tool to accelerate the discovery of synthetic lethal interactions for potential therapeutic value;it is an ongoing project in the laboratory Kolodner.


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