90891 - Biosensors and Advanced Bioanalytical Methods

Learning outcomes

At the end of the course, the student: - acquires the fundamental notions related to the use of biological molecules as reagents, - knows the basic principles of detection techniques and of the various analytical formats used in bioanalytical techniques, - knows the main types of biosensors and of bioanalytical methods and their applications, - is able to select the most suitable bioanalytical technique to solve a real problem.

Course contents

Program

 The course consists of 3 credits of lectures and 1 credit of laboratory work.

a) Lectures (3 credits, 24 hours)

1.  Introduction to biosensors: history, definition, and general classification of biosensors using molecular recognition elements and transduction systems, catalytic biosensors, and affinity-based biosensors.
2. Molecular recognition elements: enzymes, antibodies, nanobodies, non-antibody binding proteins, molecularly imprinted polymers, aptamers, DNA, RNA, peptide nucleic acids (PNAs) and cells, phage display, nanozymes.
3. Overview of the main transduction systems: biosensors with optical, electrochemical, piezoelectric, and surface plasmon resonance detection. Calorimetric biosensors.
4. Enzymatic analytical techniques: enzymes and kinetics, quantitative analysis using kinetic and endpoint methods. Simple and coupled enzymatic reactions. Detection techniques used in enzymatic analyses. Immobilization techniques. Examples of enzymatic biosensors.
5. Immunological techniques: antibodies, production of monoclonal and polyclonal antibodies, synthetic antibodies. Tracer-free and tracer-based immunological methods. Classification of homogeneous and heterogeneous immunological methods. Antibody immobilization techniques. Competitive and noncompetitive immunological methods. Cross-reactivity.
6. Lateral Flow Immunoassay (LFI) systems: basic and manufacturing components. Competitive and noncompetitive, direct and indirect LFI systems. Materials and labels used in LFI construction. Portable detectors for LFI. Examples of LFI biosensors.
7. Gene analysis techniques: DNA structure and characteristics, DNA synthesis and synthetic analogues. Gel electrophoresis, hybridization reactions (Southern blotting, filter or membrane hybridization, solution hybridization, in situ hybridization). Direct and indirect labeling. Polymerase chain reaction (PCR). Quantitative PCR by scalar dilution, with an external standard, and competitive PCR using an internal standard. Real-time PCR, Taqman probes, molecular beacons. Probe arrays (microarrays).
8. DNA nanotechnologies: Structural and functional DNA nanotechnologies. DNA origami. DNA switches, DNA tweezers, and DNA walkers. Genosensors and aptasensors. Biosensors based on DNA hybridization, label-based and label-free techniques. Examples of DNA biosensors.
9. Whole-cell biosensors: Definition and characteristics of microbial and mammalian cell-based biosensors. Reporter gene technology and examples of reporter genes. Classification of cellular bioreporters (non-specific, semi-specific, and specific). Multiplex systems. Cellular biosensors for studying protein-protein interactions: technologies based on Resonance Energy Transfer (RET) and Split technology (protein-fragment complementation assay (PCA). Iimmobilization techniques used for microbial and mammalian cells. Examples of whole-cell biosensors.
   

Review of recent scientific publications related to the topics covered in the course and applications in the medical, pharmaceutical, environmental, food, and industrial fields.

 

b) Laboratory (1 credit, 12 hours)

Readings/Bibliography

PowerPoint slides of room lectures and PDFs of the scientific publications discussed during the course are available on Insegnamenti online (http://iol.unibo.it/, access with credentials reserved for students enrolled at the University of Bologna).

Recommended texts:

Bioanalytical Chemistry (English Edition) 2nd Edition, Wiley by Susan R. Mikkelsen, Eduardo Cortón, ISBN: 1118302540, 2016 (*)

Biosensors and Nanotechnology: Applications in Health Care Diagnostics. Zeynep Altintas (Editor) ISBN: 978-1-119-06501-2 December 2017 (°)

Scientific publications will be provided.

 

(*) Recommended: Contains some useful sections for passing the exam.

(°) Not required for passing the exam, but useful for further study.

 

Teaching methods

The course consists of 24 hours of lectures and 12 hours of laboratory work.

The course materials are available online only if authorized by the University or the Department for emergency situations.

It is essential to use the materials used in the classroom during lectures and available online before the start of the course, with corresponding notification to students via the Avvisi di Virtuale tool.

During lectures (delivered via PowerPoint presentations), the course topics will be presented and discussed. Recent scientific publications related to the topics covered will also be reviewed and discussed during the course.

The laboratory exercises are intended to enable each student to acquire the necessary manual skills and knowledge of the analytical techniques covered to operate in a laboratory according to quality and safety principles. Students will process the results obtained in the laboratory using the statistical tools learned in the fundamental analytical chemistry courses and will submit a written report on the laboratory experiments.

 

It is recommended to attend classes for a better understanding of the topics.

Assessment methods

To take the exam, registration through Almaesami is required, subject to strict deadlines. If you are not interested in taking the exam, please unsubscribe. Cancellations must be made before the list closes.

To successfully pass the final exam, assessment of knowledge of the entire course content will be conducted through the following tests:

1. Presentation and discussion of a recent scientific publication chosen by the student and relevant to the topics covered in the course (max 15 minutes).
2. Oral exam on the topics covered during the course, aimed at assessing the student's acquisition of the expected knowledge and skills. Specifically, the following will be assessed: knowledge of the basic principles of bioanalytical methods, knowledge of the main types of biosensors and bioanalytical techniques, knowledge of the practical applications of the methods described above, and the ability to select the most suitable bioanalytical technique to solve a real-world problem.
   

The candidate will be examined for a total of 30 minutes (including the presentation). Assessment will be based on demonstrating critical understanding of the topics and the ability to discuss them.

 

Grading Criteria:

Points 18-24: Knowledge of a limited number of topics covered in class, some serious errors. Analytical skills emerge only with the teacher's assistance.

Points 25-29: Knowledge of a large number of topics covered in class, ability to make independent critical analysis choices, mastery of specific terminology, some minor errors and/or some flaws in presentation.

Points 30-30L: Excellent to outstanding understanding of the subject matter, thorough presentation, full mastery of specific terminology, and ability to argue.

 

Students with learning disorders and\or temporary or permanent disabilities: please, contact the office responsible (https://site.unibo.it/studenti-con-disabilita-e-dsa/en/for-students ) as soon as possible so that they can propose acceptable adjustments. The request for adaptation must be submitted in advance (15 days before the exam date) to the lecturer, who will assess the appropriateness of the adjustments, taking into account the teaching objectives.

Teaching tools

PC with screen and projector, Windows PowerPoint slides. Teaching materials (PowerPoint presentations, PDFs of publications reviewed during the course) will be made available to students at virtuale.unibo.it. Microsoft Teams platform for any online teaching options offered by the University or Department.

 

 

Office hours

See the website of Maria Maddalena Calabretta