Application of Isotopes and Antibodies in Biology and Medicine

The main objective of the course is understanding basic methods and laboratory techniques connected with isotopes and antibodies application. During the course basic aspects of nuclear physics and isotopes production will be presented. Following the course students should be familiar with principles of analytical methods in which isotopes and antibodies are applied, i.e. radioimmunoassay (RIA), radioreceptorassay (RRA), blood flow, hormone kinetics, immunocytochemistry, ELISA etc. The possibilities of these method application in biology, animal science and medicine will be presented.

Lectures (15 h):

Introduction – discussion concerning the basic problems of nuclear physics (radioactivity, dose of radioactivity, isotopes etc.).

Application of labeled substances in vivo and in vitro (hormone kinetics, blood flow through the tissue, hormone uptake by the tissue, cell proliferation).

Overview basic concepts of immunology: antigen, antibody, characteristic antigen-antibody reaction; review of methods using isotopes and/or antibodies in laboratory diagnosis. Mono- and polyclonal antibodies - the characteristics and method of their production.

Immunochemical methods (immunoassay techniques, the method ABC, fluorescent and chemiluminescent methods).

The use of antibodies in selected techniques, Part I: immunocytochemistry.

The use of antibodies in selected techniques, Part II: ELISA, Western blot, immunoprecipitation, immuno-PCR, EMSA.

Radioimmunoassay (RIA) - the principle of the method, cross-reactivity of antibodies, the test parallelism and recovery.

Principles of radioreceptorassay (RRA) – kinetics of radioligand assays, the Scatchard’s plot. Application of the RRA method in biology, medicine and pharmacology

Labs (20 h):

Immunocytochemistry (the location of apoptotic or proliferating cells on paraffin sections of tissue).

Determination of hormones and proteins by ELISA, using the ELISA method in laboratory diagnosis; determination of TSH levels in the human blood plasma

Radioimmunoassay (RIA) - determination of titer and cross-reactivity of antibodies, the assessment of antigen-antibody affinity; determination iodothyronine concentrations in animal and human blood plasma.

Determination of kinetic parameters using radioactive hormone thyroxine (biological half-life, area of distribution, metabolic clearance).

Rothfeld B.: Nuclear medicine in vitro, J.B. Lippincott Company, London, Mexico City, New York, St. Louis, Sao Paulo, Sydney, 1983.

R.V. Lloyd, Morphology methods, Cell and Molecular Biology Techniques, Humana Press, Totowa, New Jersay, 2001.

Ed Harlow, David P Lane: Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York, 1988.

Knowledge:

The student describes and defines the basic concepts and issues of nuclear physics: atom, isotope, radioactive decay, types of radiation, the measurement of radiation, radioactive series, and has knowledge of the use of radioactive isotopes in biology and medicine

Student explains the use of isotopically labeled inorganic and organic compounds in the in vivo and in vitro studies.

Student explains the importance of the most important concepts of immunology of antigen-antibody interactions, explains the ways and methods of producing antibodies mono-and polyclonal antibodies and identifies ways to use these antibodies in analytical methods.

He/she describes and characterizes the basic laboratory methods and techniques using different radioactive isotopes and antibodies, and indicates their application in biology and medicine.

Skills:The student applies radioactive isotopes and antibodies for in in vitro and in vivo experiments; can perform some analysis using isotopically labeled substances and/or antibodies

The student defines titer, affinity, and cross-reactivity of antibodies. He/she uses a radioimmunoassay (RIA) for determination of plasma concentration of hormones in animals and humans

The student applies the method of immunocytochemistry in research and diagnostics of cells and tissues. He/she interprets the results of immunocytochemical analysis.

She/he uses the ELISA method for determining the concentration of proteins and hormones.

Social competences:

The student is aware of the importance of ethical principles in carrying out animal experiments, laboratory analyzes, and the correct interpretation of the test results.

The student is aware of the responsibilities, risks and effects of radioactive substances in analytics

He/she can work in a team and manage a small team performing laboratory analyzes.

Lectures: Time limited written exam.

Labs: Demonstration of practical skills.