Leeuwenhoek discovered microorganisms and Koch established their role in infectious disease. Early diagnostic methods included microscopy and culture, but not all microbes could be observed or cultured. Later, immunological methods using vaccines and the discovery of antibodies provided defenses against pathogens. Monoclonal antibody technology was developed using hybridoma cells to produce antibodies that target specific epitopes, overcoming prior issues and allowing continuous production of effective antibodies for disease diagnosis.

2. History of Diagnostic Procedures:
Leeuwenhoek = Microorganism
Robert Koch = Infectious disease = Microorganism
1) Examination under Microscope
2) In vitro Culturing
Problems: All microbes can not be differentiated
and some can not be observed
Problems: Not all Microbes can be cultured and
beside it is very Tedious and slow process

3. 3) Immunological Diagnostic Procedures:
Edward Jenner = First vaccine = Small pox vaccine
Louis Pauster = Vaccine against Cholera
Shows Some Defensive Mechanism Involved
Which is composed Complex Cellular System
Lymphatic System

4. B.Cell “Antibody”
One B.cell = One Type of Antibody
Epitope Pathogen = Antigen
One Antigen may = More than One Epitope
“Polyclonal Antibody”:
Obtained by inserting antigen
into the body of animal
Problems
No Continuous Source
Effectiveness varies
Solution:
Culture B.Cells and
produce antibodies
through them
Problem: B.Cells can
not be cultured

5. Monoclonal Antibodies:
Antibodies Produced by Same B.Cells
Specificity Towards A Particular Epitope
Solution to Last Problem “Hydridoma Tech”
Hybrid CellB.Cell
Produces the
Desired antibody
Compatible CellPerform the
Function of
Division Myeloma Cells No Functional HGRPT gene

6. B.Cell Suspension Myeloma Cell Suspension
Combined Cell Suspension PEG
HAT media
Fused Myeloma-B.Cell Grow (other dies)
Screen those Cell Which
Produce Desired Antibodies

8. •These Monoclonal antibodies produced are used
for various purposes and regarding this
presentation, the most relevant use of Monoclonal
antibodies is in Diagnosis of diseases.