98996 - BIOCONJUGATE TECHNIQUES

Learning outcomes

At the end of the course the student has acquired knowledge of the general strategies of synthesis, purification and characterization of bioconjugates and radical reactions that lead to the oxidation of organic and biological materials, the main mechanisms of antioxidant action and the relationship between structure and antioxidant activity.

Course contents

The course will provide an extensive introduction to the mostly used bioconjugation strategies. It begins by reviewing the fundamental principles of bioconjugation including the major types of conjugates and their components, then discusses the best strategies and designs for making an optimal conjugate, and finally examines the major application areas of how bioconjugates are being used today.

At the end of the course the students will know the general strategies of bioconjugation and will have laboratory experience of synthesis, purification and characterization of bioconjugates, with reference to:

- active groups of various chemical reactions and targets of the functional groups;

- linkers and cleavable reagent systems;

- methods of synthesis, purification and characterization of the bioconjugates;

- application and research contexts in which to synthesize and use the bioconjugates.

The course is organized in two teaching modules.

Theoretical part (3 CFU)

1. Biomolecular functional targets. Overview of the functional groups of biomolecules (amino acids, peptides, proteins; polysaccharides; nucleic acids, and polynucleotides) that are most often used in making bioconjugates. The biological activity and structural properties of the biomolecules are considered as well as the location and reactivities of various functional groups that can be used for conjugation purposes.

2. Bioconjugation of micro- and nano-particles and surfaces. The types of particles being used for bioconjugation as well as the functional groups or reactive groups on their surface that exist or can be created on them for immobilizing biomolecules will be presented. Methods for particle activation, modification, and coupling, including a discussion of the reagents commonly used for these purposes will be addressed. Non-covalent approaches will be also discussed. The application fields of these bioconjugates will be reviewed.

3. The chemistry of reactive groups in bioconjugation. The major reactions used in bioconjugation techniques will be introduced. Each reaction is briefly discussed and illustrated, while providing key references from the recent literature. These reactions are further organized by the general reactivity or functional group that is involved in the associated conjugation process, such as amine, thiol, carboxylate, hydroxyl, aldehyde and ketone. Photochemical and cycloaddition reactions will be also introduced.

4. Bioconjugate Reagents. The reagents of modification and conjugation will be categorized according to structural type, reactivity, and use. Generalized protocols will be illustrated for each reagent’s most likely application. Zero-length, homobifunctional (identical reactive groups on both ends) and heterobifunctional (different reactive groups on both ends) crosslinking reagents will be exhaustively presented, comparing pros and cons of each class for specific applications.
Chemoselective and bioorthogonal reactions will be presented.

Laboratory part (2CFU)

1. Lectures dealing with

- the main techniques for bioconjugates/nanobioconjugates purification (i.e. dialysis, centrifugation, chromatography)

- the main techniques for bioconjugates/nanobioconjugates characterization (i.e. electrophoretic techniques, spectroscopic and microscopic analysis)

- presentation and discussion of the experimental work that will be carried out by the students in the lab

2. Execution of experiments in the lab during which the students will perform the synthesis purification and characterization of bioconjugates/nanobioconjugates.

Readings/Bibliography

The basic reading material will be the teachers' slides and notes; this material will be available on-line.

For further information, the following bibliography is suggested:

"Bioconjugate Techniques”, G.T.Hermanson, Academic Press, Elsevier, Amsterdam, 3nd Ed., 2013.

Teaching methods

The course consists of lectures during which will be presented and discussed

- all the topics reported in the contents section

- the experimental reactions that will be carried out by the students in the lab

The course also consists of the execution of experiments in the lab during which the students will perform the synthesis purification and characterization of bioconjugates/nanobioconjugates.

At the end of the lab activity each student will have to draw up the reports on the exercises performed (1 report for each experience), which will be delivered to the teacher.

As concerns the teaching methods of this course unit, all students must attend Module 1, 2  online, while Module 3 on health and safety is to be attended in class or on Microsoft Teams according to the modality chosen by the teacher. Information about Module 3 attendance schedule is available on the website of your degree programme.

 

 

Assessment methods

The final examination aims to ascertain the student's acquired skills in the two modules, and consists of an oral examination (30 minutes).

The course 98996 - BIOCONJUGATE TECHNIQUES together with the course 98997 - PHYSICAL ORGANIC CHEMISTRY give rise to the integrated course of 98995 - BIOCONJUGATION AND RADICAL CHEMISTRY.

The assessment of learning takes place through a final exam for each course, which ensures the acquisition of the expected knowledge and skills. The final score that is recorded for the integrated course is the arithmetic average of the 2 scores obtained in the 2 exams.

Teaching tools

The lessons are presented to students with the aid of multimedia supports. The slides are available for the students through the course web page of the Bologna University (Virtuale).

During the lab experiments, the students are requested to provide for the personal lab coat. All other personal protective equipments, all the lab equipments and all the materials necessary for the execution of the experiences are provided by the teacher.

Each student is required to sign a declaration concerning safety and health at work, which certifies that the student has received the required training to work in the lab.

Each student is also provided of a copy of the experimental procedure (already explained in the classroom) of the experience to be performed in the lab. The procedure is briefly discussed, highlighting the critical issues, before starting to operate.

Office hours

See the website of Matteo Calvaresi

See the website of Matteo Di Giosia