The document discusses various immunological techniques, including gel immunodiffusion, radioimmunoassay (RIA), fluorescence immunoassay (FIA), and single radial immunodiffusion assay (SRID). Each method is described in terms of its principles, key steps, applications, advantages, and disadvantages, highlighting their roles in detecting antigens and antibodies in clinical diagnostics and research. Despite variations in sensitivity and complexity, these techniques remain essential tools in immunology.

1. Techniques in
Immunology
Dr. Vividha Raunekar
Dr. Vividha Raunekar

2. Dr. Vividha
Raunekar
The Gel Immunodiffusion Assay, also known as the Ouchterlony Double
Diffusion Assay, is a technique used to detect the interaction between an
antigen and its specific antibody. It is commonly employed in immunology to test
for the presence of specific antibodies or antigens in a sample. The principle is
based on the diffusion of antigen and antibody through a semi-solid medium like
agar gel. When they meet, they form a visible precipitate.
Steps Involved in Gel Immunodiffusion:
1.Preparation of the Gel:
1. A thin layer of agar or agarose gel is poured into a petri dish or glass slide.
2. Wells are cut into the gel, typically arranged in a circular or linear
pattern.
2.Loading the Wells:
1. Antigen(s) and antibody are added into separate wells.
2. The antigen and antibody diffuse through the gel matrix towards each
other.
3.Diffusion Process:
1. Over time, both antigen and antibody molecules diffuse through the gel.
2. If the antigen and antibody are specific for each other, they will form a
complex in the area where they meet in the right proportions (zone of
equivalence).

3. Dr. Vividha
Raunekar
1.Precipitation Reaction:
1. At the zone of equivalence, where the optimal ratio of antigen to antibody is reached,
an antigen-antibody complex precipitates and forms a visible line or arc in the gel.
2. The precipitate line is an indication of a positive reaction, meaning the antigen and
antibody have reacted with each other.
2.Interpretation:
1. The number, location, and pattern of the precipitin lines provide information about the
antigen-antibody relationship.
2. Multiple lines may appear if multiple antigen-antibody interactions occur, or if the
antigen contains multiple epitopes that react with different antibodies.
Applications:
•Diagnosis of infectious diseases: Detects specific antibodies or antigens related to infections
(e.g., fungal infections such as Aspergillus).
•Immunology research: Used to study antigen-antibody interactions and determine cross-
reactivity.
•Vaccine testing: Helps in the analysis of immune responses.
Advantages: Simple and cost-effective.
Useful for detecting multiple antigens or antibodies in a sample.
Disadvantages:
It is a relatively slow process (takes hours to days for results).
Low sensitivity compared to more modern techniques like ELISA.
The technique is still used in certain diagnostic settings, especially for identifying fungal
infections and in veterinary immunology.

4. Dr. Vividha
Raunekar
Radioimmunoassay (RIA) is a highly sensitive and specific laboratory technique used to
measure the concentration of antigens (such as hormones, drugs, or proteins) in a
sample, based on the use of antibodies and radioactively labeled substances. It was
first developed in the 1960s by Rosalyn Yalow and Solomon Berson, and it revolutionized
clinical diagnostics, particularly in endocrinology.
Principle of Radioimmunoassay:
The principle behind RIA involves competitive binding, where a known quantity of
radioactively labeled antigen (the "tracer") competes with the unlabeled antigen in the
sample for binding to a limited amount of specific antibody. The amount of radioactivity
bound to the antibody inversely correlates with the concentration of the antigen in the
sample.
Key Steps in Radioimmunoassay:
1.Preparation of the Reaction Mixture:
1. Antibody: A specific antibody is selected for its ability to bind the antigen of
interest.
2. Radioactive Antigen (Tracer): The antigen to be measured is labeled with a
radioactive isotope, typically iodine-125 (¹² I).
⁵
3. Unlabeled Antigen: The sample (which contains the antigen to be measured) is
mixed with the radioactive antigen and antibody.

5. Dr. Vividha
Raunekar
•Competition for Antibody Binding:
•The unlabeled antigen (from the sample) and the radioactively labeled antigen
compete for a limited number of binding sites on the antibody.
•As the concentration of unlabeled antigen increases, more of it binds to the antibody,
displacing the labeled antigen.
•Separation of Bound and Free Antigens:
•After the antigen-antibody reaction reaches equilibrium, the bound antigen-antibody
complexes need to be separated from the unbound (free) antigens.
•Methods such as precipitation, centrifugation, or solid-phase techniques (e.g., using
a secondary antibody) are used to separate the bound and free fractions.
•Measurement of Radioactivity:
•The amount of radioactivity in the bound fraction (or sometimes the free fraction) is
measured using a gamma counter.
•The radioactivity in the bound fraction is inversely proportional to the concentration
of the unlabeled antigen in the sample. Lower radioactivity indicates higher antigen
concentration.
•Data Analysis:
•A standard curve is constructed using known concentrations of the antigen. The
sample’s antigen concentration is determined by comparing its radioactivity
measurement to the standard curve

6. Dr. Vividha
Raunekar
Components of RIA:
•Antigen: The substance to be measured (e.g., hormone, drug).
•Antibody: A specific antibody that binds to the antigen.
•Radioactive Label: A radioisotope (often iodine-125) is covalently attached to the
antigen.
•Gamma Counter: A device used to detect and quantify the radioactivity.
Applications of Radioimmunoassay:
1.Clinical Diagnostics:
1. Measurement of hormone levels (e.g., insulin, thyroid hormones, cortisol, growth
hormone).
2. Drug testing (e.g., detecting therapeutic drug levels or drug abuse).
3. Detecting viral antigens or antibodies (e.g., hepatitis markers).
2.Endocrinology:
1. RIA has been critical in advancing endocrinology by enabling the precise
measurement of circulating hormone levels in the blood at very low
concentrations.
3.Pharmaceutical Research:
1. Used in pharmacokinetics to study drug distribution and metabolism.
4.Biomedical Research:
1. Investigates molecular interactions and binding affinities.

7. Dr. Vividha
Raunekar
Advantages:
•High Sensitivity: Can detect very small amounts of antigen (as low as picograms).
•Specificity: The use of highly specific antibodies ensures minimal cross-reactivity
with other substances.
•Quantitative: Provides an accurate measure of antigen concentration.
Disadvantages:
•Radiation Hazard: The use of radioactive materials requires special handling,
disposal, and regulatory compliance.
•Short Shelf-Life: Radioactive isotopes have limited half-lives, meaning they lose
effectiveness over time.
•Expense: Requires specialized equipment like gamma counters, as well as
radiation safety protocols.
Alternatives to RIA:
Due to safety concerns, other non-radioactive immunoassay methods, such as
Enzyme-Linked Immunosorbent Assay (ELISA) or chemiluminescent
immunoassays, are now more commonly used. These techniques offer similar
sensitivity without the use of radioactive substances.
Despite these limitations, RIA remains a cornerstone in hormone measurement and
other sensitive antigen detection methods.

8. Dr. Vividha
Raunekar
Fluorescence Immunoassay (FIA) is a biochemical technique used to detect and
quantify specific molecules (antigens) in a sample using antibodies tagged with
fluorescent dyes. It is widely employed in diagnostics, clinical laboratories, and
research due to its high sensitivity, specificity, and ability to measure low
concentrations of antigens. The assay is based on the antigen-antibody interaction,
and the fluorescence emitted by the labeled antibodies is used as a signal to
determine the presence or concentration of the antigen.
Principle of Fluorescence Immunoassay:
FIA relies on the labeling of antibodies (or antigens) with a fluorescent molecule
known as a fluorophore. When the fluorophore is excited by a light source of a
specific wavelength, it emits light at a different, longer wavelength. The emitted
light (fluorescence) can be detected and measured, and its intensity is proportional to
the amount of antigen or antibody present in the sample.
Steps in Fluorescence Immunoassay:
1.Coating of the Antibody or Antigen:
1. A specific antibody (if the antigen is to be measured) or antigen (if the
antibody is to be measured) is immobilized on a solid surface, like a microtiter
plate or a slide.
2.Addition of Sample:
1. The sample containing the target antigen (or antibody) is added to the
reaction chamber. The antigen in the sample binds to the antibody on the solid
surface.

9. Dr. Vividha
Raunekar
•Addition of Fluorescent-Labeled Antibody:
•A secondary antibody that is specific to the antigen of interest is conjugated to a
fluorescent dye (such as fluorescein isothiocyanate [FITC], rhodamine, or Alexa Fluor).
•This fluorescent-labeled antibody binds to the antigen that is already bound to the
primary antibody (in the case of a sandwich assay), or it competes with the antigen for
binding (in the case of a competitive assay).
•Washing Step:
•After incubation, unbound antibodies are washed away. Only the bound, fluorescent-
labeled antibodies remain on the surface.
•Fluorescence Detection:
•The fluorophore is excited with a specific wavelength of light (using a fluorometer, flow
cytometer, or fluorescence microscope), causing it to emit light at a different wavelength.
•The emitted fluorescence is then measured. The intensity of the fluorescence is directly
proportional to the amount of antigen (or antibody) present in the sample.

10. Dr. Vividha
Raunekar
Types of Fluorescence Immunoassays:
1.Direct Fluorescence Immunoassay:
1. In this method, the primary antibody is directly conjugated to the
fluorophore, which binds directly to the antigen in the sample.
2. It is a simpler assay but is limited in its amplification capability.
2.Indirect Fluorescence Immunoassay:
1. A two-step process where an unlabeled primary antibody binds to the
antigen, and a secondary antibody, conjugated to a fluorophore, binds to the
primary antibody.
2. This method amplifies the signal because multiple secondary antibodies can
bind to a single primary antibody, increasing the fluorescent signal.
3.Sandwich Fluorescence Immunoassay:
1. This method involves two antibodies: one that captures the antigen and
another, fluorescently labeled, that detects it.
2. The antigen is "sandwiched" between the capture antibody and the detection
antibody. This assay is highly specific and sensitive.
Competitive Fluorescence Immunoassay:
In this format, a fixed amount of labeled antigen competes with the unlabeled
antigen (from the sample) for binding to a limited number of antibody sites.
As the concentration of the antigen in the sample increases, less labeled antigen
binds to the antibody, leading to reduced fluorescence.

11. Dr. Vividha
Raunekar
Fluorescent Labels (Fluorophores):
Common fluorophores used in FIA include:
•Fluorescein Isothiocyanate (FITC): Emits green fluorescence when excited at 490 nm.
•Rhodamine: Emits red fluorescence.
•Alexa Fluor dyes: A family of fluorophores with a wide range of excitation and
emission wavelengths.
•Phycoerythrin (PE): A red-orange fluorophore used in flow cytometry.
•Cyanine (Cy3, Cy5): Used for applications requiring longer emission wavelengths.
Applications of Fluorescence Immunoassay:
1.Clinical Diagnostics:
1. Measurement of hormones, tumor markers, viral antigens, or antibodies in serum
or plasma.
2. Detection of infectious diseases (e.g., HIV, hepatitis).
3. Allergy testing by measuring specific IgE antibodies.
2.Pharmaceutical Research:
1. Screening for drug compounds or therapeutic antibodies.
2. Studying molecular interactions.
3.Environmental Monitoring:
1. Detecting contaminants or toxins in water, food, or environmental samples.
4.Flow Cytometry:
1. Fluorescence immunoassays are integrated into flow cytometry for analyzing
cells and particles by measuring surface or intracellular antigens.

12. Dr. Vividha
Raunekar
Advantages of Fluorescence Immunoassay:
•High Sensitivity: Fluorescent labels can detect very low concentrations of antigens
(as low as femtomolar levels).
•Specificity: The use of highly specific antibodies reduces cross-reactivity and
improves accuracy.
•Non-Radioactive: Unlike radioimmunoassays, FIA uses non-hazardous fluorescent
labels, making it safer to use in laboratories.
•Quantitative: The fluorescence intensity can be measured and correlated to the
concentration of the antigen or antibody in the sample.
Disadvantages of Fluorescence Immunoassay:
•Photobleaching: Fluorophores can degrade over time when exposed to light,
reducing the intensity of the fluorescence signal.
•Interference: Background fluorescence from the sample matrix or the plasticware
can interfere with the measurement.
•Equipment Cost: Fluorescence detection requires specialized equipment like
fluorometers or fluorescence microscopes, which may be expensive.
Conclusion:
Fluorescence immunoassay is a powerful tool for detecting and quantifying
biological molecules with high sensitivity and specificity. Its use of non-radioactive
fluorescent labels makes it safer than other methods like radioimmunoassay, and its
versatility has made it an indispensable technique in diagnostics, research, and
industry.

13. Dr. Vividha
Raunekar
Single Radial Immunodiffusion Assay (SRID), also known as the Mancini method, is a
quantitative immunological technique used to measure the concentration of antigens
(such as proteins) in a sample. It is widely used for measuring immunoglobulins (IgG,
IgA, etc.) and other proteins, especially in clinical settings.
Principle of Single Radial Immunodiffusion:
SRID is based on the diffusion of an antigen through an agar gel that contains a
uniformly distributed specific antibody. As the antigen diffuses radially from a central
well into the antibody-containing gel, it forms antigen-antibody complexes. When an
optimal ratio (zone of equivalence) is reached, a visible precipitin ring forms around
the well. The diameter of the ring is directly proportional to the concentration of the
antigen in the well.

14. Dr. Vividha
Raunekar
Steps Involved in Single Radial Immunodiffusion:
• Preparation of the Gel:
• Agarose gel is mixed with a specific antibody that is directed against the antigen of interest.
• The gel is poured into a petri dish or on a glass slide and allowed to solidify.
• Wells Cutting:
1. Small wells are punched into the gel using a template or well-cutter, typically arranged in a grid
pattern.
•Addition of the Antigen:
•The sample containing the antigen (along with standards of known antigen concentration) is placed into the
wells.
•The antigen diffuses radially outward from the well into the antibody-containing gel.
•Formation of the Precipitin Ring:
•As the antigen diffuses, it reacts with the antibody in the gel, forming insoluble antigen-antibody complexes.
•A visible ring of precipitate forms at the point where the antigen and antibody concentrations are optimal (zone
of equivalence).
•The larger the amount of antigen, the further it diffuses, leading to a larger precipitin ring.
•Measurement:
•After a specific incubation time (typically 24–72 hours), the diameter of the precipitin ring is measured.
•The diameter of the ring is directly proportional to the concentration of the antigen in the well.
•Data Analysis:
•A standard curve is generated by plotting the ring diameters of known antigen concentrations against their
respective concentrations.
•The concentration of antigen in the unknown sample is then determined by comparing its ring diameter to the
standard curve.

15. Dr. Vividha
Raunekar
Components of Single Radial Immunodiffusion:
•Agarose Gel: The gel provides a medium through which the antigen diffuses.
Agarose is preferred because of its transparency and consistency.
•Antibody: The specific antibody against the antigen of interest is mixed
uniformly into the gel.
•Antigen: The substance to be quantified is placed into the wells.
•Standards: Known concentrations of the antigen are run alongside the unknown
sample to create a standard curve for quantification.
Applications of Single Radial Immunodiffusion:
1.Clinical Diagnostics:
1. Measuring serum proteins such as immunoglobulins (IgG, IgA, IgM).
2. Quantifying complement proteins (C3, C4) and other plasma proteins.
3. Evaluating protein deficiencies and monitoring immune disorders (e.g.,
hypogammaglobulinemia).
2.Vaccine Quality Control:
1. SRID is used to measure the concentration of viral antigens in vaccines,
particularly in the production and standardization of influenza vaccines.
3.Research:
1. Protein quantification in various biological samples such as blood, tissue
homogenates, and cell cultures.
4.Animal Health Diagnostics:
1. Measurement of antibodies or antigens in veterinary diagnostics.

16. Dr. Vividha
Raunekar
Advantages of Single Radial Immunodiffusion:
•Quantitative: SRID provides an accurate measurement of antigen concentration
in the sample.
•Simple to Perform: It does not require sophisticated equipment or technical
expertise, making it accessible for routine use.
•Stable Reaction: The antigen-antibody complexes are stable and produce a
clear precipitin ring, allowing for easy measurement.
Disadvantages of Single Radial Immunodiffusion:
•Time-Consuming: The diffusion and precipitin ring formation can take 24 to 72
hours to complete, which is slower than other immunoassays like ELISA.
•Limited Sensitivity: SRID is less sensitive than other methods, such as enzyme-
linked immunosorbent assays (ELISA) or radioimmunoassay (RIA). It is best suited
for medium to high concentrations of antigen.
•Not Suitable for Complex Mixtures: The assay works well when there is only
one antigen-antibody interaction. Complex mixtures may produce overlapping
precipitin rings, making interpretation difficult.

17. Dr. Vividha
Raunekar
Factors Affecting SRID:
1.Antibody Concentration: The concentration of antibody in the gel must be optimized
to ensure proper ring formation. Too much or too little antibody can distort the results.
2.Gel Thickness: A uniform thickness of the gel is essential for consistent diffusion and
accurate measurements.
3.Incubation Time: Longer incubation allows for better diffusion, but excessive
incubation can lead to indistinct or overly large rings.
4.Temperature: The assay should be conducted at an appropriate temperature (usually
room temperature) to ensure proper antigen-antibody interaction and diffusion.
Conclusion:
Single Radial Immunodiffusion is a classical immunoassay method for quantifying
antigen concentrations in biological samples. While newer techniques like ELISA have
largely replaced SRID in many applications due to their faster and more sensitive results,
SRID remains valuable in specific clinical and research settings, particularly for
measuring immunoglobulins and proteins in vaccines.

18. Dr. Vividha
Raunekar
Immunodiagnostics
Immunodiagnostics refers to a collection of diagnostic techniques that leverage
the specificity of antigen-antibody reactions to detect and measure substances in
the body, such as pathogens, proteins, or abnormal cells. These tests are widely
used in clinical laboratories for diagnosing infections, autoimmune diseases,
cancers, allergies, and more.
Key Principles of Immunodiagnostics:
Immunodiagnostics is based on the fundamental concept of antigen-antibody
binding. When an antigen (such as a protein, pathogen, or marker molecule)
binds to a specific antibody, the formation of this complex can be detected using
various techniques, providing information about the presence or quantity of the
antigen.
Types of Immunodiagnostic Techniques:
1.Enzyme-Linked Immunosorbent Assay (ELISA):
1. One of the most widely used immunoassays.
2. Detects the presence of antigens or antibodies in a sample by using
enzyme-labeled antibodies. Upon binding, a substrate is added, leading to
a detectable color change.

19. Dr. Vividha
Raunekar
•Radioimmunoassay (RIA):
•Uses radioactive isotopes attached to antibodies or antigens. The amount of radioactivity
correlates with the amount of antigen-antibody binding, allowing for quantification of the target
molecule.
•Immunofluorescence:
•Fluorescent-labeled antibodies bind to specific antigens. The emitted fluorescence can be
detected using a microscope or fluorometer.
•Used in detecting viruses, bacteria, or other pathogens in tissues.
•Western Blotting:
•Separates proteins by size using gel electrophoresis, followed by identification with specific
antibodies.
•Commonly used in the detection of specific proteins in research or disease markers like HIV
proteins.
•Lateral Flow Assays (LFAs):
•Also known as rapid tests (e.g., pregnancy tests or COVID-19 antigen tests).
•A sample migrates across a membrane strip with antibodies to produce a visual result
(typically in the form of lines).
•Flow Cytometry:
•Measures the physical and chemical characteristics of cells or particles.
•Uses fluorescently labeled antibodies to detect specific markers on the surface or inside cells.
Commonly used in immunophenotyping, such as in leukemia diagnostics.
•Agglutination Tests:
•Based on the clumping of particles when an antigen-antibody interaction occurs.
•Used in blood typing and pathogen detection.

20. Dr. Vividha
Raunekar
Applications of Immunodiagnostics:
1.Infectious Diseases: Detecting pathogens like bacteria, viruses, fungi, and
parasites through their antigens or antibodies produced in response to infection
(e.g., HIV, hepatitis, influenza).
2.Cancer Detection: Identifying tumor markers (such as PSA in prostate cancer
or HER2 in breast cancer) using specific antibodies.
3.Autoimmune Disorders: Diagnosing autoimmune diseases like rheumatoid
arthritis, lupus, or celiac disease by detecting autoantibodies.
4.Allergy Testing: Identifying specific IgE antibodies related to allergens.
5.Hormone and Drug Monitoring: Measuring hormone levels (e.g., insulin,
thyroid hormones) or therapeutic drug levels using specific antibodies.
Advantages of Immunodiagnostics:
•High specificity: Antibody-antigen binding is highly specific, reducing false
positives.
•Sensitivity: Able to detect even small quantities of antigens or antibodies.
•Versatility: Can be used to detect a wide range of diseases and biomarkers.
Limitations:
•False Positives/Negatives: If not carefully designed, cross-reactivity or
technical errors can lead to inaccurate results.
•Cost and Equipment: Some immunodiagnostic methods require sophisticated
equipment and are expensive.

21. Dr. Vividha
Raunekar
Immunotherapy
Immunotherapy is a therapeutic approach that uses the body’s immune system to fight diseases, particularly
cancer, autoimmune diseases, and chronic infections. The goal of immunotherapy is to enhance, suppress, or
modify the immune response to achieve a therapeutic effect.
Types of Immunotherapy:
1.Cancer Immunotherapy: Cancer immunotherapy works by stimulating the immune system to recognize and
attack cancer cells more effectively. Several types include:
1. Checkpoint Inhibitors:
1.These drugs block proteins (e.g., PD-1, CTLA-4) on immune cells or cancer cells that inhibit the
immune response. By blocking these checkpoints, immune cells can recognize and kill cancer cells.
2.Examples: Pembrolizumab (Keytruda), Nivolumab (Opdivo).
2. CAR T-Cell Therapy:
1.Involves genetically modifying a patient’s T-cells to express a receptor (chimeric antigen receptor,
CAR) that targets cancer cells.
2.Once reintroduced into the patient’s body, these CAR T-cells can recognize and destroy cancer cells.
3.Used in blood cancers like leukemia and lymphoma.
3. Cancer Vaccines:
1.Cancer vaccines help train the immune system to recognize cancer-related antigens and trigger an
immune response.
2.Example: Sipuleucel-T (Provenge) for prostate cancer.
4. Cytokine Therapy:
1.Cytokines like interferons and interleukins are used to boost the immune system’s response to
cancer.
2.These can activate immune cells or enhance the production of other immune-boosting molecules.

22. Dr. Vividha
Raunekar
•Allergy Immunotherapy:
•Involves controlled exposure to allergens to desensitize the immune system over time. This can reduce allergic
reactions in conditions like hay fever, asthma, or food allergies.
•Methods include allergy shots or sublingual immunotherapy (placing small doses of the allergen under the
tongue).
•Autoimmune Disease Immunotherapy:
•Immunotherapy can be used to suppress the abnormal immune response in autoimmune diseases, where the
immune system mistakenly attacks the body’s own tissues.
•Examples include monoclonal antibodies (e.g., Rituximab) that target specific immune cells involved in
autoimmune conditions like rheumatoid arthritis, lupus, or multiple sclerosis.
•Monoclonal Antibody Therapy:
•Monoclonal antibodies (mAbs) are lab-made antibodies designed to bind specifically to a target antigen (such as
a protein on cancer cells).
•Some monoclonal antibodies can deliver cytotoxic agents directly to cancer cells, while others mark the cells for
destruction by the immune system.
•Example: Trastuzumab (Herceptin) for HER2-positive breast cancer.
•Immune Modulators:
•These therapies alter the immune response, either enhancing it in cases of cancer and infections or suppressing
it in cases of autoimmune diseases.
•Drugs like Thalidomide or Lenalidomide are used in cancers like multiple myeloma to enhance the immune
response against cancer cells.
•Adoptive Cell Transfer (ACT):
•A form of immunotherapy where a patient’s T-cells are collected, modified, expanded in the lab, and then
reintroduced into the patient’s body to fight cancer cells.

23. Dr. Vividha
Raunekar
Applications of Immunotherapy:
1.Cancer Treatment:
1. Immunotherapy is especially transformative in treating cancers like melanoma, lung
cancer, and hematological cancers (e.g., leukemia, lymphoma).
2.Autoimmune Diseases:
1. Used in managing rheumatoid arthritis, lupus, psoriasis, and Crohn's disease by
suppressing the hyperactive immune response.
3.Chronic Infections:
1. Immunotherapy is being explored for chronic infections like HIV or hepatitis, where
boosting the immune response could help control or eliminate the virus.
Advantages of Immunotherapy:
•Targeted Treatment: Immunotherapy often targets specific molecules or cells, reducing
damage to healthy tissue compared to traditional therapies like chemotherapy or radiation.
•Durability: In some cases, immunotherapy can offer long-lasting protection by "teaching"
the immune system to remember and attack cancer cells or pathogens in the future.
Limitations:
•Side Effects: Immunotherapy can lead to immune-related adverse effects, such as
inflammation, autoimmune reactions, and systemic side effects (e.g., fever, fatigue).
•Response Variability: Not all patients respond to immunotherapy, and it may only be
effective for certain types of cancers or diseases.
•Cost: Immunotherapies, especially CAR T-cell and monoclonal antibody therapies, can be
extremely expensive and may not be accessible to all patients.

24. Dr. Vividha
Raunekar
Thank you