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Posted On: 03/23/2025 11:25:43 AM
Post# of 155794
If people don’t want chatGPT questions posted I won’t share them. But I see it as a source of information and have been using it in other ways beside here. I know the accuracy isn’t 100% but simple questions seem reasonable to be… that’s my disclaimer…
I asked a basic question: how are monoclonal antibodies made… FYI
Monoclonal antibodies (mAbs) are made using a laboratory process that produces identical, specific antibodies designed to target a particular antigen (such as a protein on the surface of a virus, cancer cell, or other pathogen). Here’s a step-by-step overview of how they are made:
1. Antigen Identification and Immunization
• Scientists first identify the specific antigen they want the mAbs to target.
• A lab animal, usually a mouse, is injected with the antigen to stimulate its immune system.
• The animal produces a variety of B cells, each generating a unique antibody against the antigen.
2. B Cell Isolation
• After the immune response is triggered, B cells are harvested from the animal’s spleen.
• These B cells produce antibodies but cannot survive long outside the body.
3. Hybridoma Formation
• To enable long-term antibody production, the B cells are fused with myeloma cells (cancerous plasma cells) in vitro.
• This fusion creates hybridoma cells, which have two key properties:
• They produce a single (monoclonal) antibody.
• They can divide indefinitely, allowing for continuous antibody production.
4. Selection and Screening
• The hybridoma cells are cultured in a selective medium that allows only the successfully fused cells to survive.
• The hybridomas are screened to identify the ones that produce the desired antibody with high specificity and affinity for the target antigen.
5. Cloning and Expansion
• The selected hybridoma cells are cloned to ensure uniform antibody production.
• These cells are grown in large-scale bioreactors or cell culture systems to produce large quantities of the monoclonal antibodies.
6. Purification
• The antibodies are purified from the culture medium using processes like protein A affinity chromatography.
• The purified mAbs are further refined to remove impurities and ensure consistency.
7. Modification and Humanization
• Since mAbs from mice can trigger an immune response in humans, they are often genetically engineered to make them more compatible with humans.
• This process is called humanization, where the mouse-derived antibody sequences are replaced with human sequences, except for the antigen-binding sites.
• Some mAbs are fully human, produced using transgenic mice or phage display technology.
8. Quality Control and Testing
• The final mAbs undergo rigorous quality control testing to confirm their purity, potency, and safety before being approved for clinical use.
✅ Production Methods
• Traditional hybridoma method: Still widely used for many mAbs.
• Recombinant DNA technology: Increasingly common for producing fully human mAbs by using engineered cell lines (e.g., Chinese hamster ovary [CHO] cells).
This process allows for the large-scale production of highly specific monoclonal antibodies, which are used in treatments for cancer, autoimmune diseases, infectious diseases, and more.
I asked a basic question: how are monoclonal antibodies made… FYI
Monoclonal antibodies (mAbs) are made using a laboratory process that produces identical, specific antibodies designed to target a particular antigen (such as a protein on the surface of a virus, cancer cell, or other pathogen). Here’s a step-by-step overview of how they are made:
1. Antigen Identification and Immunization
• Scientists first identify the specific antigen they want the mAbs to target.
• A lab animal, usually a mouse, is injected with the antigen to stimulate its immune system.
• The animal produces a variety of B cells, each generating a unique antibody against the antigen.
2. B Cell Isolation
• After the immune response is triggered, B cells are harvested from the animal’s spleen.
• These B cells produce antibodies but cannot survive long outside the body.
3. Hybridoma Formation
• To enable long-term antibody production, the B cells are fused with myeloma cells (cancerous plasma cells) in vitro.
• This fusion creates hybridoma cells, which have two key properties:
• They produce a single (monoclonal) antibody.
• They can divide indefinitely, allowing for continuous antibody production.
4. Selection and Screening
• The hybridoma cells are cultured in a selective medium that allows only the successfully fused cells to survive.
• The hybridomas are screened to identify the ones that produce the desired antibody with high specificity and affinity for the target antigen.
5. Cloning and Expansion
• The selected hybridoma cells are cloned to ensure uniform antibody production.
• These cells are grown in large-scale bioreactors or cell culture systems to produce large quantities of the monoclonal antibodies.
6. Purification
• The antibodies are purified from the culture medium using processes like protein A affinity chromatography.
• The purified mAbs are further refined to remove impurities and ensure consistency.
7. Modification and Humanization
• Since mAbs from mice can trigger an immune response in humans, they are often genetically engineered to make them more compatible with humans.
• This process is called humanization, where the mouse-derived antibody sequences are replaced with human sequences, except for the antigen-binding sites.
• Some mAbs are fully human, produced using transgenic mice or phage display technology.
8. Quality Control and Testing
• The final mAbs undergo rigorous quality control testing to confirm their purity, potency, and safety before being approved for clinical use.
✅ Production Methods
• Traditional hybridoma method: Still widely used for many mAbs.
• Recombinant DNA technology: Increasingly common for producing fully human mAbs by using engineered cell lines (e.g., Chinese hamster ovary [CHO] cells).
This process allows for the large-scale production of highly specific monoclonal antibodies, which are used in treatments for cancer, autoimmune diseases, infectious diseases, and more.
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