91309 - BIOCHIMICA APPLICATA MEDICA

Course Unit Page

Academic Year 2021/2022

Learning outcomes

At the end of the course, the student will have learned:
- the main biochemical techniques and methodologies of biochemical analysis
- the rationale behind the applications of these technologies to medicine and the clinic;
- how to apply biochemical methods to the clinical analyses, to understand and correctly interpret the results also from a diagnostic/prognostic point of view;
- communicate to patients the correct reading of reports of simple biochemical-clinical analyses.

Course contents


1. UV-VIS Absorption Spectroscopy

a) Physical principles of absorption of electromagnetic radiation
b) Lambert-Beer law
c) Instrumentation and applications

Fluorescence Spectroscopy

a) Physical principles of the phenomenon
b) Instrumentation and applications


3. Chemiluminescence

4. Principles of cytofluorimetry
a) Instrumentation and applications

5. Liquid Chromatography
a) gel-filtration
b) ion exchange
c) affinity

6. Centrifugal sedimentation techniques

7. Electrophoresis

a) Principles of the technique
b) SDS-PAGE
c) Isolectrofocusing
d) 2D gel-electrophoresis

8. Immunochemistry

a) Antibody (Ab) structure and interaction with antigen (Ag)
b) Primary and secondary immune response
c) Polyclonal and monoclonal Ab production (Mab)
d) Precipitation of the Ab-Ag complex (equivalence zone)
e) Simple and double immunodiffusion techniques
f) Immunoelectrophoresis
g) ELISA, Immunofluorescence, DELFIA, RIA
h) Physiological and pathological aspects

9. Molecular Biology Techniques

a) Restriction enzymes and DNA analysis
b) Hybridization and transfer techniques on cellulose supports (blotting)
c) Fingerprinting and Polymorphisms
d) PCR, principles and applications

Laboratory Practice:


1.Assay of protein content in biological samples: Bradford method-.


2. Assay of enzyme activity, construction of a Michaelis-Menten curve and calculation of KM and Vmax


3. Determination of the antioxidant power of compounds


4. Cell counting and staining methods of cellular structures.




Readings/Bibliography

  • Metodologie biochimiche e biomolecolari. Mauro Maccarone Ed. Zanichelli
  • Biochimica Clinica. Luigi Spandrio Edizioni Sorbona
  • Biochimica Applicata, autori Stoppini-Bellotti Editore EdiSES, Napoli
  • Metodologia biochimica - Le bioscienze e le biotecnologie in laboratorio A cura di Keith Wilson e John Walker Edizioni Raffaello Cortina Editore, Milano

Teaching methods

The course includes 4 frontal credits, in which a series of biochemical techniques and their medical application will be considered, and 2 laboratory credits in which problems such as: determination of the protein content of a solution, determination of enzymatic kinetic parameters, cell counting or staining of cellular samples will be addressed from a practical point of view. The presentations of the lectures will be deposited in the Virtual Learning Platform.

 

“As concerns the teaching methods of this course unit, all students must attend Module 1, 2 [https://www.unibo.it/en/services-and-opportunities/health-and-assistance/health-and-safety/online-course-on-health-and-safety-in-study-and-internship-areas] online, while Module 3 on health and safety is to be attended in class. Information about Module 3 attendance schedule is available on the website of your degree programme”

Assessment methods

In order to take the exam, it is necessary the registration on the almaesami website, respecting the deadlines. Those who are not able to enroll by the due date, are required to communicate promptly (and in any case before the official closure of the registration lists) the problem to the teacher by e-mail.The learning assessment will be done through the final oral exam. The student will discuss a short elaborate concerning the application of one of the techniques covered during the course and general questions about the techniques considered during the course. The evaluation will be based on the answers given to each question and the ability demonstrated to have learned the fundamental notions to achieve the educational objectives. The definition of that knowledge and its critical mastery will contribute to the final grade. The student will also be required to submit a report on the laboratory, which will be evaluated.

Teaching tools

Virtuale platform.

Office hours

See the website of Christian Bergamini