Applied Biochemistry

A.Y. 2021/2022
8
Max ECTS
72
Overall hours
SSD
BIO/10
Language
Italian
Learning objectives
The course aims to provide students with the theoretical (56 hours of frontal lesson) and practical (16 hours of laboratory practice) basis of the most advanced techniques used in biomedical research and their main applications.
The aim of the course is to provide students with the basic notions for an analytical approach to the different problems encountered in both basic and applied research laboratories.
The training objective of the course is to present and critically discuss biochemical methods to highlight potentiality, limits and complementarity. The contents of the course are constantly updated to train professionals who can fit into a biomedical laboratory with a theoretical preparation suitable for the practical use of the most modern methodologies.
Expected learning outcomes
The students' learning outcomes are evaluated by the teacher through a written examination of applied biochemistry learning based on the program carried out during the course.
The examination of the students by the teacher will be mainly aimed at assessing the theoretical and practical knowledge and understanding of biochemical techniques and the assessment of the skills acquired by the student at the completion of the course itself. In particular, the student's ability to use the language and the skills acquired to solve problems typical of biochemical laboratories with exercises of cases present in the literature will be evaluated. Finally, upon completion of the laboratory part, each student will have to present a report on the practical activities carried out and demonstrate the capacity to prepare a report and to critically discuss the results obtained.
Course syllabus and organization

Linea AK

Responsible
Lesson period
First semester
More specific information on the delivery modes of training activities for academic year 2021/2022 will be provided over the coming months, based on the evolution of the public health situation.
Course syllabus
The following topics will be explained during the course:
UV-Visible Spectrophotometry: Recalls on the nature of electromagnetic radiation. UV-VIS spectrophotometry (principles, instrumentation and applications). Absorption spectra. Lambert-Beer's law and its quantitative applications, the concept of calibration curve. Spectrophotometric dosage of protein concentration. Application of spectrophotometry in enzymatic dosages
Chromatographic techniques for the purification, separation, identification and quantitative analysis of low (metabolites, drugs, hormones, etc.) and high (nucleic acids and proteins) molecular weight molecules: General principles. About TLC. Ion exchange chromatography. Exclusion chromatography, Affinity chromatography. HPLC and GC (stationary phases, detectors and main applications).
Electrophoretic techniques: General principles. Zonal electrophoresis on cellulose acetate; serum proteins. Gel electrophoresis: agarose, polyacrylamide gel electrophoresis (PAGE and SDS-PAGE). Isoelectrofocalization (IEF). Two-dimensional electrophoresis. Detection methods and quantitative evaluations (gel and blotting stains). Capillary electrophoresis.
Mass spectrometry techniques for the qualitative and quantitative analysis of low (metabolites, drugs, hormones, etc.) and high (nucleic acids and proteins) molecular weight molecules : General principles and instrumentation (sources and analyzers).
Techniques for the study of proteins: methods for sequencing, determination of secondary and tertiary structure, and post-translational changes and mutations. Proteomics background
Molecular biology techniques: basic concepts of molecular biology. Recombinant DNA (vectors, restriction enzymes and other enzymes, plasmid DNA purification, obtaining and screening of libraries). DNA sequencing. Methods for the study of gene transcription and expression (mRNA levels, transcription activity of promoters). Methods for the study of the interaction between macromolecules (DNA-proteins, protein-proteins). Polymerase chain reaction (PCR) and real time PCR: principles and applications.
Methods of labeling biological molecules: Redioactive labeling. Energy and speed of radioactive decay. Detection and measurement of radioactivity by scintillation. Autoradiography. Labeling with non-radioactive systems.
Cell cultures and techniques for cellular fractionation
Immunochemical and immunometric techniques: structure of antibodies and on the antigen-antibody reaction. Free phase and gel immunoprecipitation. Immunodiffusion and agglutination assays. Radioimmunological and enzymatic immunoassays, turbidimetric and nephelometric methods.
Exercises in a teaching laboratory
The course includes mandatory practical laboratory exercises on some of the topics covered in the course and includes an individual report.

Materiale di riferimento in italiano
Nel portale Ariel sono disponibili un eserciziario che raccoglie i temi di esame più recenti e le risoluzioni di alcuni problemi.
Possono essere consultati i seguenti testi:
Metodologie di base per le scienze biomolecolari - Reed R, Holmes D, Weyers J, Jones A - Zanichelli ed., ultima edizione
Metodologie di base per la biochimica e la biotecnologia - Ninfa AJ, Ballou DP - Zanichelli ed., ultima edizione
I principi di biochimica di Lehninger - Nelson & Cox - Zanichelli ed., ultima edizione
Prerequisites for admission
To take the exam, students must have passed Biochemistry and Organic Chemistry I exams
Teaching methods
The course includes 56 hours of lectures and 16 hours of laboratory practices. Attendance at the laboratory exercises is compulsory for admission to the exam; a written report on the laboratory activities carried out is required.
Teaching Resources
he iconographic material shown during the lessons can be downloaded from the Ariel portal.
The following texts can be consulted:
Basic Methodologies for Biomolecular Sciences - Reed R, Holmes D, Weyers J, Jones A - Zanichelli ed., last edition
Basic Methodologies for Biochemistry and Biotechnology - Ninfa AJ, Ballou DP - Zanichelli ed., last edition.
The principles of biochemistry of Lehninger - Nelson & Cox - Zanichelli ed., last edition.
Assessment methods and Criteria
The students' learning outcomes are evaluated by the teacher through an oral examination of applied biochemistry learning based on the program carried out during the course.
The examination of the students by the teacher will be mainly aimed at assessing the theoretical and practical knowledge and understanding of biochemical techniques and the assessment of the skills acquired by the student at the completion of the course itself. In particular, the student's ability to use the language and the skills acquired to solve problems typical of biochemical laboratories with exercises of cases present in the literature will be evaluated. Finally, upon completion of the laboratory part, each student will have to present a report on the practical activities carried out and demonstrate the capacity to prepare a report and to critically discuss the results obtained
BIO/10 - BIOCHEMISTRY - University credits: 8
Practicals: 16 hours
Lessons: 56 hours
Professor: Caruso Donatella

Linea LZ

Responsible
Lesson period
First semester
Teaching methods.
If the situation allows it, lessons will be held in the classroom. Each lesson will be video-recorded and made available asynchronously through the Ariel platform. If the situation worsens or there are no classrooms suitable to contain all the students of the course respecting the social distancing, the lessons will be delivered through the Microsoft Teams platform and can be followed synchronously according to the scheduled time and asynchronously because recorded through the same platform. The laboratory practices will be held for 25% in the presence of a teaching laboratory suitably equipped for the Covid emergency and for the remaining 75% in a virtual laboratory through specially developed videos. If this is not possible, all the laboratory practices will be carried out in the virtual laboratory. Please note that, regardless of the mode of delivery, all laboratory practices are mandatory.

Program and reference material.
The program and reference material of the lectures will not be changed. All supporting didactic material will always be made available through the Ariel platform.

Methods of learning verification and evaluation criteria.
If the situation allows it and classrooms are available to contain the students who want to take the exam while maintaining the social distancing, the exam will be held in written form according to the modalities reported in the section "Examination Modalities". Alternatively, the examination will take place in oral form using the Microsoft Teams platform, Skype or similar.
Course syllabus
The following topics will be explained during the course:
UV-Visible Spectrophotometry: Recalls on the nature of electromagnetic radiation. UV-VIS spectrophotometry (principles, instrumentation and applications). Absorption spectra. Lambert-Beer's law and its quantitative applications, the concept of calibration curve. Spectrophotometric dosage of protein concentration. Application of spectrophotometry in enzymatic dosages
Chromatographic techniques for the purification, separation, identification and quantitative analysis of low (metabolites, drugs, hormones, etc.) and high (nucleic acids and proteins) molecular weight molecules: General principles. About TLC. Ion exchange chromatography. Exclusion chromatography, Affinity chromatography. HPLC and GC (stationary phases, detectors and main applications).
Electrophoretic techniques: General principles. Zonal electrophoresis on cellulose acetate; serum proteins. Gel electrophoresis: agarose, polyacrylamide gel electrophoresis (PAGE and SDS-PAGE). Isoelectrofocalization (IEF). Two-dimensional electrophoresis. Detection methods and quantitative evaluations (gel and blotting stains). Capillary electrophoresis.
Mass spectrometry techniques for the qualitative and quantitative analysis of low (metabolites, drugs, hormones, etc.) and high (nucleic acids and proteins) molecular weight molecules : General principles and instrumentation (sources and analyzers).
Techniques for the study of proteins: methods for sequencing, determination of secondary and tertiary structure, and post-translational changes and mutations. Proteomics background
Molecular biology techniques: basic concepts of molecular biology. Recombinant DNA (vectors, restriction enzymes and other enzymes, plasmid DNA purification, obtaining and screening of libraries). DNA sequencing. Methods for the study of gene transcription and expression (mRNA levels, transcription activity of promoters). Methods for the study of the interaction between macromolecules (DNA-proteins, protein-proteins). Polymerase chain reaction (PCR) and real time PCR: principles and applications.
Methods of labeling biological molecules: Redioactive labeling. Energy and speed of radioactive decay. Detection and measurement of radioactivity by scintillation. Autoradiography. Labeling with non-radioactive systems.
Cell cultures and techniques for cellular fractionation
Immunochemical and immunometric techniques: structure of antibodies and on the antigen-antibody reaction. Free phase and gel immunoprecipitation. Immunodiffusion and agglutination assays. Radioimmunological and enzymatic immunoassays, turbidimetric and nephelometric methods.
Exercises in a teaching laboratory
The course includes mandatory practical laboratory exercises on some of the topics covered in the course and includes an individual report.
Prerequisites for admission
To take the exam, students must have passed the Biochemistry and Organic Chemistry I exams.
Teaching methods
The course includes 56 hours of lectures and 16 hours of laboratory practices. Attendance at the laboratory exercises is compulsory for admission to the exam; a written report on the laboratory activities carried out is required.
During the course some lessons will be dedicated to the resolution of examination texts.
Teaching Resources
The iconographic material shown during the lessons can be downloaded from the Ariel portal.
In the Ariel portal there is an exercise book that collects the most recent exam themes and the solutions to some problems.
The following texts can be consulted:
Basic Methodologies for Biomolecular Sciences - Reed R, Holmes D, Weyers J, Jones A - Zanichelli ed., last edition
Basic Methodologies for Biochemistry and Biotechnology - Ninfa AJ, Ballou DP - Zanichelli ed., last edition.
The principles of biochemistry of Lehninger - Nelson & Cox - Zanichelli ed., last edition.
Assessment methods and Criteria
The students' learning outcomes are evaluated by the teacher through a written examination of applied biochemistry learning based on the program carried out during the course. The examination of the students by the teacher will be mainly aimed at assessing the theoretical and practical knowledge and understanding of biochemical techniques and the assessment of the skills acquired by the student at the completion of the course itself. In particular, the student's ability to use the language and the skills acquired to solve problems typical of biochemical laboratories with exercises of cases present in the literature will be evaluated. Finally, upon completion of the laboratory part, each student will have to present a report on the practical activities carried out and demonstrate the capacity to prepare a report and to critically discuss the results obtained.
BIO/10 - BIOCHEMISTRY - University credits: 8
Practicals: 16 hours
Lessons: 56 hours
Professor: Mitro Nico