Production technology: hybridoma generation
To produce monoclonal antibodies on an industrial and medical scale, a special biotechnological procedure is used—hybridoma generation. This process combines the best features of the natural immune response and tumor cells.
The technological steps include:
- Cell fusion. Immune B-lymphocytes (e.g., mouse splenocytes) that produce the required antibodies are taken as starting material and fused with myeloma cells. Myeloma cells are tumorous and "immortal," but they are incapable of producing their own antibodies.
- Use of medium. The fusion process of these two cell types is triggered in the presence of a specific agent: polyethylene glycol.
- Hybridoma formation. As a result, a hybrid cell is formed. From the B-lymphocyte, it inherits strict antigen specificity, and from the myeloma, the unique ability to divide indefinitely.
Thus, a single hybridoma can produce homogeneous, highly specific antibodies indefinitely.
Cultivation methods
Following the successful creation of hybridoma cells, the task arises to mass-produce them to obtain a sufficient quantity of antibodies. There are two main approaches to cultivation:
- In vitro — growing cells outside a living organism in special artificial cell cultures with nutrient media.
- In vivo — injecting the hybridoma into the peritoneal cavity of a laboratory mouse. In this case, monoclonal antibodies accumulate in high concentrations in the animal's ascetic fluid.
Antibodies obtained in this manner are actively used for clinical diagnostics, as well as for targeted therapy of oncological and severe autoimmune diseases.
Modified monoclonal antibodies
Since antibodies are initially obtained using animal cells (mouse lymphocytes), their direct administration to humans can cause adverse reactions. To avoid this, biotechnologies allow the creation of modified variants widely used in endocrinology, the treatment of autoimmune diseases, and transplantology.
Chimeric monoclonal antibodies Represent a combination of mouse and human protein components.
- Structure: The variable region (the so-called Fab fragment), responsible for antigen specificity, is taken from mouse antibodies. The remaining constant part of the molecule is replaced with a human IgG fragment.
- Application: Effective in treating autoimmune pathologies such as rheumatoid arthritis.
Humanized monoclonal antibodies These drugs are structurally as close as possible to human immunoglobulins and contain a minimal amount of animal protein.
- Structure: Only small gene segments of the hypervariable regions (CDRs) derived from rats are precisely "embedded" into human immunoglobulin genes.
- Application: Actively used in transplantology to prevent the rejection of transplanted organs.
Striking examples of humanized antibodies are daclizumab and basiliximab—drugs that act as interleukin-2 (IL-2) receptor blockers, thereby preventing the unwanted activation of T-lymphocytes.
Briefly: answers to topical questions
What are some examples of chimeric monoclonal antibody drugs?
Chimeric monoclonal antibody drugs include rituximab and cetuximab. Their International Nonproprietary Names use the suffix -ximab.
- Rituximab — a drug containing variable regions of mouse protein and constant regions of human protein.
- Cetuximab — a drug whose structure is a combination of mouse and human components.
By what methods are fully human monoclonal antibodies obtained?
Fully human monoclonal antibodies are produced using genetic engineering methods. These drugs belong to the group of genetically engineered biological products.
To create an artificial protein that reproduces the minimal part of an antibody with the antigen-binding site, phage display technology is used.
- Phage display technology — a method for creating and selecting antigen-binding molecules of desired specificity from combinatorial B-lymphocyte gene libraries using bacteriophages.
To remember the essence of a hybridoma, use the formula: "Specificity from the lymphocyte + Immortality from the myeloma = Hybridoma (antibody factory)."
Class and exam questions
Why is polyethylene glycol used in antibody production technology?
Polyethylene glycol is added to the medium to stimulate and facilitate the physical fusion process of immune B-lymphocytes with myeloma cells.
How do chimeric antibodies differ from humanized ones?
In chimeric antibodies, an entire Fab fragment (variable region) is taken from an animal (mouse). In humanized antibodies, there is even less animal protein—only rat segments of the hypervariable regions (CDRs) are embedded into the human immunoglobulin.
What role do daclizumab and basiliximab play?
They are humanized monoclonal antibodies that block IL-2 receptors. This prevents T-lymphocyte activation and protects against transplant rejection.
What else to study on this topic
- Mechanisms of monoclonal antibody binding to antigen epitopes
- Application of antibodies in clinical laboratory diagnostics
- Oncological therapy using hybridoma technologies
- The role of IL-2 receptor blockers in transplantology
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