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Lonely Musafir

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  • Deepak Sharma
  • October 08, 2019 08:18:06 AM

A Little About Us

Lonelymusafir writes blogs and vlogs on his Himalayan Biking Trips to Spiti, Sikkim, Laddakh. Lonelymusafir's blogs Ayodhya Pahar, Prurliya an offbeat tribal(Santhali) location are rare to find. Landscape and portrait Photography along with urdu shayari and sad poetry makes this blog a versatile one. The blogs is practical and based on personal experience sharing. The blog contains detailed travelogues and itinerary on Trichy, Pudducherry, Angriya Cruise. Do's and Don'ts for Bikers taking Royal Enfield Rides to Himalayas, based on personal Experiences is rare to find.

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Sariska Bike Ride

A short bike ride to Sariska National Park, Alwar, Rajasthan with RF Riders

A short bike ride to Sariska National Park, Alwar, Rajasthan

An Old Age Man

Portrait of an Old Age Man from Rurals of Alwar, Rajasthan.

An Old Age Man from rurals of Alwar, Rajasthan

The Cure for Corona- Understanding Monoclonal Antibodies

Monoclonal Antibodies are the cure for Corona Virus. They are produced using HAT medium using Hybridoma Technology. This article explains in very simple terms/ Language the process of mAB production.

Corona (COVID-19) disease has been declared a pandemic by World Health Organisation, with 1,42,755 cases as per the latest data and claiming more than 5000 lives till 13.03.2020 midnight.

Researchers across the globe are working on the cure for COVID-19. By the time they come out with a cure, we the common man need to understand the cure.

I being a Graduate in Biotechnology trying to make an attempt to make general masses understand as how a cure is developed for such a pandemic.

What are Monoclonal Antibodies?

The Monoclonal Antibodies (mAB or moAb) are our fighting force against the virus. These are antibodies specific for one type of Antigen (Here- Corona Virus 2019-nCoV).

Monoclonal Antibody- Antigen Specific Antibody

Antigen is an extracellular (Foreign Cell) species which activates our immune system (i.e. Stimulates Antibody Production) in our body . It may be a bacteria, virus, protozoa or fungi etc.

How will the mAB fight Corona Virus 2019-nCoV?

In case of COVID-19 the Corona Virus 2019-nCoV is the antigen. An antigen after entering a host body (Human is our case) stimulates B-Lymphocytes (Immunity Cells) which is then converted into a Plasma cell. These plasma cells are the fighters which starts secreting Antibodies for the antigens sensed by B-Lymphocytes. These antibodies kill the Antigen by blocking its active sites.

Antigen-B-Cell reaction and formation of Plasma Cell for antibody generation.

How can we produce Monoclonal Antibodies for Corona Virus?

There is a set procedure of producing mAB for any antigen. Let me explain it in simple steps.

  1. Inject the antigen in a living body.
  2. Isolate Spleen Cells from the living body.
  3. Fuse these spleen cells with a Cancerous (Myeloma) Cell for form Hybridomas.
  4. Culture these Hybridomas in selective (HAT) Media.
  5. Isolate best quality Monoclonal Antibodies from the viable cells.

Let us now understand the process in detail and in simple terms.

Why Inject Virus in living body?

We need to produce Antibodies for a deadly antigen like corona virus 2019-nCoV. Researchers will first inject corona virus 2019-nCoV to a mice or a guinea pig.

The injection will be done intravenously or in simple words virus will be directly injected into the blood stream of that mice.

This will activate the immune response in the mice and later antibodies will be generated and that is what we want.

Why Spleen Cells?

As the injection has been done directly into the blood stream, the antigens will be trapped into the spleen, which is the secondary lymphoid organ.

Virus Injection & the Immune response inside the mice spleen

The virus 2019-nCoV, will stimulate the B-Cell (immune response) inside the Spleen cells converting it into plasma cells generating Antibodies to kill Corona Virus 2019-nCoV.

Why fuse the Spleen cells with Cancerous cells?

After taking out the mice spleen one can ask why it is required to fuse the spleen cells with myeloma cells? Why can’t we simply extract the antibodies from plasma cells using a culture medium?

The reason why plasma cells can not be used to extract Antibodies directly is due to their shorter life spans. The plasma cells have a very short life span which is not enough for the process of cell culture and antibody extraction.

Fusion of Myeloma and Plasma Cells

Myeloma cells are cancerous cells which are immortal cells and have very fast rate of multiplication. Hence fusing a plasma cell with Myeloma cell will result in immortalisation of Plasma cells by formation of a Hybridoma (Hybrid cell of Plasma and Myeloma).

So now we have a cell which has an endless life and potential to generate Antibodies.

Why we need to Culture Hybridomas in selective Media?

During the fusion of Myeloma and Plasma cells five types of cells can possibly form.

Five possible type of Cells that can result after fusion of Myeloma and Plasma cells using Polyethyleneglycol

Out of which Only Fused Plasma (Plasma+Plasma), Hybridoma (Plasma+Myeloma) and Unfused Plasma (Single Plasma) cells are of our use as they only have property of generating 2019-nCoV specific antibodies.

HAT medium- How it selects the desirable cells?

HAT stands for Hypoxanthine, Aminopterin and Thymidine. These are the three ingredients of the selective medium chosen to culture the Hybridomas so that it can generate desirable antibodies.

Two modes of Nucleic Acid Production- De Novo and Salvage Process

All mother cells divides into daughter cells by replication of DNA, for this syntheses of Nucleic acid is required. This synthesis of Nucleic Acid can be done by two pathways.

De-Novo pathway– Where Nucleic Acid is synthesised from basic molecules.

Salvage Pathway– Where Nucleic acid is synthesised from degraded nucleotides.

Now in HAT Media, due the presence of Aminopterin cells can not replicate thorugh De Novo Pathway, as Aminopterin blocks a key enzyme Dihydrofolatereductase.

Denovo and Salvage pathway. Use of Aminopterin in HAT medium.

Now the cells in HAT media has only way to replicate is through Salvage Process. But, here is the catch. Cells need HGPRT(Hypoxanthine Guanine Phospho Ribosil Transferase) enzyme to relplicate through Salvage process.

Only HGPRT + cells will replicate in HAT Medium

Myeloma cells- NO HGPRT

Plasma Cells(Fused and Unfused) and Hybridoma cells- Has HGPRT enzyme, Hence replicates in HAT Medium.

Plasma cells and fused plasma cells having very small life span will automatically perish in HAT media and Hybridoma cells will only survive.

The Cure

These Hybridomas will yield the Monoclonal Antibodies, which will be our weapon against the Corona Virus- 2019-nCoV.

Many researchers are working on the same lines for generating a cure for this deadly virus. Till the time they succeed, let’s keep the fingers crossed.

PC: Google Images & Sajid Shaikh

Arunachal Bike Trip Part-II (Dirang to Tawang)

LonelyMusafir Travels to Tawang from Dirang Crossing mighty Sela Pass, Frozen Sela lake, Freezed Jaswantgarh. The Detailed Video Narrates how we travelled across snowy roads and freezing wind.

LonelyMusafir Travels to Tawang from Dirang Crossing mighty Sela Pass, Frozen Sela lake, Freezed Jaswantgarh. The Detailed Video Narrates how we travelled across snowy roads and freezing wind.

Evening Indolence- An Old Lady from Santhal Tribe

An Old Lady from Santhal Tribe-Ayodhya Pahar Region, Puruliya, West Bengal, India

An Old Lady from Santhal Tribe-Ayodhya Pahar Region, Puruliya, West Bengal, India

Arunachal Bike Trip Part-1(Guwahati to Dirang)

This is a vlog of My bike trip to Arunachal Pradesh in January 2020. This is Part-1 which covers my journey from Guwahati to Dirang Via Lapool, Bhairavkunda, Kalaktanag, Shergaon, Rupa and Bomdila.

Arunachal Bike Trip Part-1 (Guwahati to Dirang) via BhairavKunda- Kalaktang- Shergaon-Rupa-Bomdila

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