Learning outcomes of the course unit
The course of Applied Biochemistry is a part of teaching or course-integrated Biochemistry-Applied Biochemistry.
The Course aims to enable the student to know and understand the general principles of experimental biochemistry, biological materials, principles and equipment used for analytical and preparative purposes including for the purpose of allowing the student the ability to use the knowledge and understanding gained to understand the significance of biochemical investigation, discipline-based research, and appropriated the principles and methods normally used in biochemical laboratory.
The student at the end of the Course, using the knowledge of Organic Chemistry previously acquired, must demonstrate knowledge and understanding about:
-material used in biochemical experiments
-preparative and analytical separation techniques
-techniques based on biospecific interactions
In addition, students, applying the knowledge and understanding acquired, shall be able,even collaborating with other professionals, to help explain:
-how to investigate the mechanism of action of a drug
-investigate as to understanding the molecular basis of diseases
-how to study the properties of an enzyme marker of a disease
-how to measure an antigen in a biological fluid complex.
The student must have sufficient technical language to explain the various stages of extraction, separation,purification, molecular and functional characterizatio of macromolecules of biochemical interest.
Basic knowledge of General Chemistry, Organic Chemistry, Cell Biology.
Course contents summary
First part:general criteria in biochemical experimentation. Biological materials. Second part:methods of separation and purification of proteins and other molecules of biochemical interest: centrifugation, salting out, dialysis, chromatography,electrophoresis. Third part: biochemical applications of visible and ultraviolet spectroscopy. Study of enzymatic activity. Immunochemical, radioisotope and radioimmunologic techniques. Potentiometric and polarographic techniques Overview of molecular biology.
General principles of biochemical experimental investigation
Methods of separation and purification of proteins and other molecules of biochemical interest: centrifugation, salt fractionation, dialysis and ultrafiltration.
Chromatographic techniques. Principles and modes of chromatography. Chromatografic performance parameters. Adsorption chromatography; ion-exchange chromatography;molecular exclusion (gel filtration) chromatography; affinity chromatography; high performance liquid chromatography (HPLC);gas-liquid chromatography.
Electrophoretic techniques. General principles. Support media and detection methods. Electrophoresis of proteins and of nucleic acids. Isoelectric focusing. Capillary electrophoresis.
Spectroscopic techniques. Properties of electromagnetic radiation and interaction with matter. Ultraviolet and visible light spectroscopy. Spectrofluorimetry. Applications in biochemical analysis.
Enzyme assay and kinetic analysis.
Immunochemical techniques. Immunoglobulin structure. Production and purification of polyclonal and monoclonal antibodies. Immunoprecipitation . Labeling antibodies. Enzyme-
Radioisotope techniques and radioimmunoassay (RIA).
Potenziometric and polarographic techniques
K.Wilson,J.Walker:''Principi e tecniche di Biochimica e Biologia molecolare''; M.C.Bonaccorsi di Patti,R.Contestabile,M.L.Di Salvo "Metodologie Biochimiche"2e.; M.Maccarrone "Metodologie biochimiche e biomolecolari"
Oral lesson frontal carried out with the help of communication tools.
Plesa refer to "Reference texts" useful for the preparation of exam.
Assessment methods and criteria
The verification of the achievement of the objectives set by the Integrated Course includes a written test with open answers with time available for 2 hours and 30 minutes. Through questions regarding the course content will be determined whether the student has achieved the goal of understanding and knowledge. The answer to four questions of wich three of Biochemistry and the fourth concerning the general principles of biochemical investigations and biological materials used, methods of separation and purification of proteins and other molecules of biochemical interest, the principles and application of spectroscopic techniques and immunochemical techniques will allow to ascertain whether the student has attained the aim of applying the acquired knowledge .Each answer will be evaluated with a score from 0 to 30/30 praise.The final evaluation will consist of adding the vote obtained in the answer to each individual question and dividing by the number of questions. To pass the exam it is necessary to have a grade higher than eighteen in Applied Biochemistry response.