PB4211 Immuno Technology Laboratory Syllabus:

PB4211 Immuno Technology Laboratory Syllabus – Anna University PG Syllabus Regulation 2021

COURSE OBJECTIVES

The course aims to,
 provide hands-on-experience on handling animal for research and various relevant I immunological techniques like ELISA, Flow cytometry etc.
 provide practical experience on performing and understanding immunoassays forevaluating drugs.
 to obtain laboratory training for different Immuno technological techniques.

LIST OF EXPERIMENTS

1.Preparation of antigen and Routes of immunization (Intraperitoneal, Sub-cutaneous, Intramuscular, Intra- nasal, Oral – VIRTUAL DEMO)
2. Methods of bleeding (Tail bleeding, Intravenous, intraorbital – VIRTUAL DEMO)
3. Collection of serum, storage and purification of total IgG (salt precipitation).
4. Evaluation of Antibody titre by direct ELISA
5. Evaluation of Antigen by Sandwich ELISA
6. Characterization of antigens by native and SDS-PAGE
7. Characterizations of antigens by Western blot analysis – Wet and semi dry transfer
8. Conjugation of Immunoglobins (Streptavidin, colloidal gold)
9. Methods for prototype development of Immunodiagnostics (ICT card)
10. Blood smear identification of leucocytes by Giemsa stain 16
11. Separation of mononuclear cells by Ficoll-Hypaque
12. Separation of splenocytes and proliferation against mitogens
13. Primer design using softwares.
14. Gene DNA amplification by random / specific primers.
15. Western Blotting
16. Gene amplification by PCR.

TOTAL: 90 PERIODS

LIST OF EQUIPMENTS REQUIRED

Microscopes
Purification columns
Microplate reader
UV spectrometer
PAGE apparatus
Western blot apparatus
Centrifuge
Haemocytometer
Cell counter
Metabolic analyser
PCR

COURSE OUTCOMES

At the end of the course, learners will be able to

• Demonstrate the immunization methods and handling of animals
• Carry out the experiment on immunological assays for identifying drugs and vaccines.
• Describe the principles of PCR and their uses in genetic engineering.
• Illustrate the principle, detection and quantification of protein by bioanalytical techniques
• Learn about the gene amplification and methods for analysis of DNA
• Apply methods adapted in gene synthesis

REFERENCES:

1. “Antibodies”, Cold Spring Harbour Laboratory, 1988.
2. Goldsby, R.A. et al.“Kuby Immunology”. 6thEdition, W.H. Freeman, 2002.
3. Turgeon, Mary Louise. “Immunology and Serology in Laboratory Medicine”, 2nd Edition, Elsevier, 2007.
4. Brostoff J et al., “Clinical Immunology”, 6th Edition, Gower Medical Publishing, 2002.
5. Coligan, J. E. et al, “Current Protocols in Immunology”, 4thEdition John Wiley & Sons, 1994.
6. Paul, “Fundamental of Immunology”, 4thEdition, Lippincott Raven, 1999.