Hybridoma Technology: Animated explanation



hello friends this is Sajid and in this video session we'll learn about the hybridoma technology so this hybridoma technology is used for the production of large number of monoclonal antibodies against a particular antigen and these monoclonal antibodies are used for the different purposes for example in the treatment of cancer and also as anti-venom for example anti snake venom and TB venom and despite a venom basically this monoclonal antibodies are produced by a white blood cell which is called as plasma cell whenever any antigen enters in the body especially the extracellular antigen the B cell or P lymphocyte get activated by it and it is converted into the plasma cell and finally this plasma cell secretes the antibodies against that particular antigen now we might think if the antigen is administered into the body of animal and that animal is going to produce plasma cell against that particular antigen which produces monoclonal antibodies against it we can isolate this plasma cell from the lab animal and they can culture into the artificial media and in artificial media this plasma cell will produce antibodies for us it is correct the plasma cells can be cultured into the artificial media but the biggest problem is that the lifespan of plasma cells is very short it is from few weeks to few months and that's why we cannot produce antibodies for a long time from culturing them into the artificial media in hybridoma technology this problem of short lifespan of plasma cell is overcome by fusing it by a cancerous cell which is called as myeloma cell myeloma cell due to it is the cancer it is immortal in nature so we have fused one cell which has the ability for production of antibodies and other cell which is immortal in nature the new cell which is produced due to the fusion of this two cell is called as the hybridoma cell and therefore this hybrid cell gains the property of both cell so this cell is going to produce antibodies as well as it is immortal in nature let us see the actual procedure of hybridoma technology first of all the antigen against which the antibodies are to be produced is injected intravenously to the lab animal this lab animal it generally mice or any other mammal since the antigen is given intravenously that is directly into the blood the antigens are trapped into the spleen spleen is the secondary lymphoid organ inside this plane the antigen interacts with the B cell and activates them to convert into the plasma cell once the plasma cells are produced they will secrete the large number of monoclonal antibodies against the antigen injector after this this plasma cell producing the monoclonal antibodies against our injector and ijen they are isolated from this plane after the purification of plasma cell it is fused with the myeloma cell the fusion procedure is carried out in the presence of polyethylene glycol and sometimes electroporation can also be used since the fusion procedure is random it is not like that the plasma cells will always fused with the myeloma cell and therefore at the end of fusion procedure five different types of cells are produced and they are fused plasma cells fused myeloma cells infused plasma infused myeloma cell and the most important hybridoma cell among these five cells we are only interested in the hybridoma cell and therefore the next step is the selection of hybridoma selection is carried out on the hat media h a T here the H is for hypo xanthine a is for aminopterin and t is for thymidine these are the contents which are present in the hat media see whenever any cell divides in the artificial media before dividing it must synthesize the new copy of the DNA and for that it requires the synthesis of nucleotides nucleotides can be synthesized by two ways either the cell can go for the salvage pathway or it can go for de novo synthesis in case of salvage pathway cells uses the degraded part of old nucleotide to produce the new nucleotide and in case of de novo pathway completely new nucleotides are synthesized using small metabolites like different sugars different amino acid which are present in the media in case of had media cells cannot operate de novo pathway and this is due to the presence of aminopterin in the media this aminopterin it blocks one key enzyme which is required to operate the de novo pathway and this enzyme is dihydrofolate reductase well this enzyme is blocked cells cannot operate the de novo pathway okay therefore there is only one option left for the cells in the hat' media and that is the salvage pathway cells can operate salvage pathway but for this also there is one condition and this condition is that say must have one enzyme which is called as a GPR D that is hypoxanthine gone in phospho ribose ill transferase and for operating the salvage pathway which has this particular enzyme they can use the hypose Anton and thymidine as precursor plasma cells how this enzyme but myeloma cells don't I want to remind you that five different cells are produced after the fusion procedure of plasma and memory cells and desire fused plasma fused myeloma hybridoma and fused plasma and infused myeloma cell among this self plasma cells have a GP RT and so the fused plasma cells myeloma cell don't have a GP RT enzyme and therefore fused myeloma will also like it what about hybridoma in case of hybridoma one cell is plasma and therefore it will have AG PRT enzyme correct now what will happen if all these five different kind of cells are transferred on the hat media cells which lack it GP RT enzyme will not grown the hat media and these cells are fused myeloma and infused myeloma the other three types of cells can grow on the hat media but lifespan of plasma and the fuse plasma is very short and within few weeks they will automatically die no need to separately kill them what about hybridoma again it is a GP RT positive and it is immortal in nature it will keep growing with the hat media it has no limit and therefore at the end hybridoma cells will survive on the hat media in this way the hybridoma is selected and used for the continuous antibody production

21 Comments

  1. the black lushai said:

    The Most well explained video for Hybridoma Technlology

    May 22, 2019
    Reply
  2. anu jha said:

    Pls pls pls make more videos sir….i already liked nd subscribed

    May 22, 2019
    Reply
  3. anu jha said:

    Oh my god…no words to express my gratitude ❤ 💕 💓 💙

    May 22, 2019
    Reply
  4. Maanvi Khairkar said:

    People who disliked this video are stupid

    May 22, 2019
    Reply
  5. Cole Mimi Ralte said:

    please make more Biotechnology videos, totally living for it!! 😊

    May 22, 2019
    Reply
  6. amer sayeed said:

    Sajid , just loved ur video , u explained soo perfectly !

    May 22, 2019
    Reply
  7. Pratibha Kolan said:

    Excellent Sajid..Very well explained😊

    May 22, 2019
    Reply
  8. Abhirami Gouri said:

    Sooo useful thnku

    May 22, 2019
    Reply
  9. Versha Bhardwaj said:

    Outstanding performance!!

    May 22, 2019
    Reply
  10. Tamanya Roy said:

    Could not be more clearee

    May 22, 2019
    Reply
  11. Suba Rasu said:

    Tqsm sir😊

    May 22, 2019
    Reply
  12. Beenish Sofi said:

    👍👍👍👍👍

    May 22, 2019
    Reply
  13. Neha Gupta said:

    Why do we use aminopterin in media?

    May 22, 2019
    Reply
  14. rtg 123 said:

    tq

    May 22, 2019
    Reply
  15. Abominatrix650 said:

    This is a highly effective video. Thank you for creating it.

    May 22, 2019
    Reply
  16. One minute laugh said:

    Clear to understand. Thanks man!

    May 22, 2019
    Reply
  17. straming with shuvranil said:

    Great information

    May 22, 2019
    Reply
  18. Yash Sagar said:

    Very good work

    May 22, 2019
    Reply
  19. BIBISHNA AV said:

    Thank you …..

    May 22, 2019
    Reply
  20. Vyankatesh Zambare said:

    Really nice explanation! And one more thing that I liked is, you used your original accent while explanation. Thanks again!

    May 22, 2019
    Reply
  21. helen paras said:

    You explained it very well.

    May 22, 2019
    Reply

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