BMB514: Immunological techniques (5 ECTS)

STADS: 01001401

Level
Bachelor course

Teaching period
The course is offered in the autumn semester.

Teacher responsible
Email: kskjoedt@health.sdu.dk

Additional teachers
ckoch@health.sdu.dk

Timetable
Show entire timetable
Show personal time table for this course.

Prerequisites:
None

Academic preconditions:
Theoretical Immunology (BMB512) must be followed.

Course introduction
To give the student practical experience in immunological techniques and the production and characterization of immunoreactants, and to provide them with a grasp of the great potential of these methods in biological research and development.

Expected learning outcome
When the course is completed, the student is expected to:

• have gained a better understanding of theoretical immunology, through "hands-on” experience of immune system
• be able to manufacture, purify and characterise antibodies produced in, for example, rabbit or mouse
• use antibodies for qualitative and quantitative analysis
• be able to develop optimise and validate immunological assays
• be able to determine qualitative parameters such as antigen specificity, epitope specificity, affinity and the involvement of the types of bonds in the reaction between antigen and antibody
• work with cell cultures in the manufacture and characterisation of monoclonal antibodies
• work in accordance to regulations in a sterile laboratory
• assess the results of tests based on the use of immunological reagents.

Subject overview
I. Production of monoclonal antibodies, including immunisation, fusion, selection, cloning and the determination of specificity.
II. Analysis of polyclonal antibodies, purification of IgG, analysis of purity and specificity and the production of mono-specific preparations from poly-specific antisera.
III. Quantitative immunological techniques with special focus on the "sandwich" format in the form of Enzyme-Linked Immunosorbent Assay (ELISA). With regard to ELISA, skills in developing and configuring the assay will be acquired and applied to the quantification of an antigen in various body fluids. The effects of interference by heterophilic anti-Ig antibodies and human anti-animal IgG antibodies will also be examined. In addition, quantitative radio-immunoassay (RIA) and quantitative immunoelectrophoresis and immunodiffusion techniques will be performed.
IV. Analytical immunological techniques. In connection with the production of monoclonal antibodies, students will be instructed about the setup and application of analytical ELISA to identify relevant hybridomas and to monitor cloning. The students are introduced to the use of analytical immunoelectrophoresis techniques to study the specificity of polyclonal antibodies and to monitor chromatographic separations. The students will also be schooled in the use of double immunodiffusion to illustrate the concepts of immunological identity, non-identity and cross-reaction. Western blots will be performed in relation to the identification of specific antigens in a biological fluid and to evaluation of the specificity of the antibodies employed. In the course of the ELISA exercises, knowledge about new technologies to define the relative epitope specificity of monoclonal antibodies will also be acquired.
V. In the course teaches students the use of fluorochrome-labelled antibodies in FACS analysis to identify specific cell types and quantification of receptors on these cells. In addition, students are trained in the use of immunohistochemistry to test antigen localization in tissue sections, using enzyme-labelled antibodies.
VI. Students will be instructed in the immunoprecipitation, isolation and characterization of radioactive-labelled protein mixtures, using specific antibodies in combination with SDS-PAGE and autoradiography. Introduction to immunoagglutination will be acquired through exercises centred on the AB0 blood group system.

Literature
There isn't any litterature for the course at the moment.

Website
This course uses e-learn (blackboard).

Prerequisites for participating in the exam
None

Assessment and marking:

1. Written report, passed/fail and internal marking by the teacher. (5 ECTS).(01001402).

Expected working hours
The teaching method is based on three phase model.

Laboratorieøvelser (10 kursusdage med hver 6 timer, ialt 60 timer).
Educational activities

Educational form
Laboratory exercises, 10 days with 6 hours per day.

Language
This course is taught in Danish.

Remarks
The course is build up with individual themes. A written manual (130 pages) will clarify the individual themes.

Course enrollment
See deadline of enrolment.

Tuition fees for single courses
See fees for single courses.