Quality Control and Analysis for Biopharmaceuticals
A.Y. 2023/2024
Learning objectives
This course will provide the theoretical basis of the main spectroscopic (IR, NMR, CD), mass spectrometric analysis techniques such as IE, CI and ESI, DSC and XRPD. It will give to the student essential tools to determine the structure of organic and bioorganic compounds. Particular importance is given to the analysis of the data and the strategy for their interpretation, also through problems solving strategies. Basic notions will be given concerning the most popular analytical techniques applied for the characterization of biopharmaceuticals, including chromatography, capillary and gel electrophoresis, mass spectrometry as well as emission and absorption UV-Vis spectroscopy. Moreover, various procedures for potency (biological activity) testing will be shown, including ligand and receptor binding assays, cell culture-based assays, biochemical assays (such as enzymatic assays) and transgenic animal-based assays.
Expected learning outcomes
At the end of the course the student is expected 1) to prove a basic knowledge of the principles of the most popular analytical techniques for the characterization of biopharmaceuticals 2) to be able to plan analytical strategies for the characterization of biopharmaceuticals; 3) to be able to understand the results obtained from instrumental analyses of biopharmaceuticals; 4) to prove a basic knowledge of main methods employed for measuring biological activity of biopharmaceuticals; 5) to learn how to handle the spectroscopic data and to use them in an integrated and comparative way with the aim to extract all the information about the structure of organic and bioorganic molecules; 6) to be able to communicate clearly and with an appropriate language the knowledge and the acquired interpretative ability of the various spectra and 7) to develop a good autonomy for the identification of the structures of organic molecules.
Lesson period: First semester
Assessment methods: Esame
Assessment result: voto verbalizzato in trentesimi
Single course
This course can be attended as a single course.
Course syllabus and organization
Single session
Responsible
Lesson period
First semester
Prerequisites for admission
Knowledge of the fundamentals of physics, chemistry, organic chemistry, and biochemistry.
Assessment methods and Criteria
For the teaching unit "Spectroscopic methods in bioorganic chemistry" the exam is a two-hours written test consisting of a theory question to assess theoretical knowledge of spectroscopic techniques illustrated during the course and an exercise concerning the spectral data interpretation of an organic molecule with known structure with the aim of assessing the student's ability to apply the knowledge and understanding acquired in analyzing spectroscopic data and to justify the logical process followed. The results of the written test, notified through the University platform, will then be than discussed in an individual interview.
For the teaching unit "Analytical and pharmacodynamic characterization of biopharmaceuticals" the exam consists of a two-hour written test for pharmacodynamics and of an oral test for analysis. In detail, the written test consists of 30 multiple choice questions (4 answers, only one of which is correct), whereas the oral test is aimed at verifying the knowledge of the fundamentals of biopharmaceutical analysis and biological activity measurement. Results of the written test are posted on the course' electronic blackboard within a few days after the test, whereas for the oral test the result will be communicated to the candidate at the end of the test. Evaluation criteria include assessment of the knowledge and ability of cogitating on the topics discussed during the teaching unit.
The final mark will consist of the CFU-weighted average of the partial marks for the modules.
For the teaching unit "Analytical and pharmacodynamic characterization of biopharmaceuticals" the exam consists of a two-hour written test for pharmacodynamics and of an oral test for analysis. In detail, the written test consists of 30 multiple choice questions (4 answers, only one of which is correct), whereas the oral test is aimed at verifying the knowledge of the fundamentals of biopharmaceutical analysis and biological activity measurement. Results of the written test are posted on the course' electronic blackboard within a few days after the test, whereas for the oral test the result will be communicated to the candidate at the end of the test. Evaluation criteria include assessment of the knowledge and ability of cogitating on the topics discussed during the teaching unit.
The final mark will consist of the CFU-weighted average of the partial marks for the modules.
Pectroscopic methods in bioorganic chemistry
Course syllabus
GENERAL PART: electromagnetic spectrum and electromagnetic radiation, characterization of electromagnetic radiation, electromagnetic radiation and interactions with matter, spectroscopic techniques.
INFRARED SPECTROSCOPY: Basic principles: frequency of infrared absorption and chemical structure.
The IR spectrum. Characteristic absorption: hydrocarbons (alkanes, alkenes, alkynes, aromatics), alcohols, ethers, halides, carbonyl compounds, amines. IR spectrometers: classic, Fourier transform IR spectrophotometer (FT-IR). Reflection techniques: ATR, DRIFT, RAIRS. Applications of IR spectroscopy.
NUCLEAR MAGNETIC RESONANCE, NMR (Nuclear Magnetic Resonance): general principles, properties of nuclei, magnetic field, and electromagnetic radiation (radiofrequency), the resonance phenomenon
Proton magnetic resonance: Chemical shift, Spin-spin coupling, Spin decoupling. Overhauser effect.
13C-NMR spectroscopy: Introduction. Carbon spectrum recording techniques. Chemical displacements. Spin 13C-1H coupling. DEPT and APT spectra. NMR correlation spectroscopy: 1H-1H COSY, 1H-13C HETCOR-HSQC-HMBC. NMR spectroscopy of other important nuclei 15N, 19F, 31P. Applications
MASS SPECTROMETRY. Introduction. Instrumentation, introduction system, sources, analyzers. Low and high-resolution molecular ion. Molecular ion recognition. Fragmentations and rearrangements.
Mass spectra of some chemical classes. CIRCULAR DICROISM: Introduction. Principles. Applications
TECHNIQUES OF CHARACTERIZATION OF THE SOLID STATE: Differential Scanning Calorimetry and X-rays difractometry. Basic principles and applications.
For all the analytical techniques, problems in the area of biomolecule characterization will be addressed.
INFRARED SPECTROSCOPY: Basic principles: frequency of infrared absorption and chemical structure.
The IR spectrum. Characteristic absorption: hydrocarbons (alkanes, alkenes, alkynes, aromatics), alcohols, ethers, halides, carbonyl compounds, amines. IR spectrometers: classic, Fourier transform IR spectrophotometer (FT-IR). Reflection techniques: ATR, DRIFT, RAIRS. Applications of IR spectroscopy.
NUCLEAR MAGNETIC RESONANCE, NMR (Nuclear Magnetic Resonance): general principles, properties of nuclei, magnetic field, and electromagnetic radiation (radiofrequency), the resonance phenomenon
Proton magnetic resonance: Chemical shift, Spin-spin coupling, Spin decoupling. Overhauser effect.
13C-NMR spectroscopy: Introduction. Carbon spectrum recording techniques. Chemical displacements. Spin 13C-1H coupling. DEPT and APT spectra. NMR correlation spectroscopy: 1H-1H COSY, 1H-13C HETCOR-HSQC-HMBC. NMR spectroscopy of other important nuclei 15N, 19F, 31P. Applications
MASS SPECTROMETRY. Introduction. Instrumentation, introduction system, sources, analyzers. Low and high-resolution molecular ion. Molecular ion recognition. Fragmentations and rearrangements.
Mass spectra of some chemical classes. CIRCULAR DICROISM: Introduction. Principles. Applications
TECHNIQUES OF CHARACTERIZATION OF THE SOLID STATE: Differential Scanning Calorimetry and X-rays difractometry. Basic principles and applications.
For all the analytical techniques, problems in the area of biomolecule characterization will be addressed.
Teaching methods
Teaching is organized in Lectures, with recommended attendance. Classroom exercises aimed at reading and understanding spectra as well as on structural confirmation of simple organic molecules will be provided during the course of lectures.
Teaching Resources
R.M. Silverstein, F.X. Webster Identificazione spettrometrica di composti organici Terza edizione-Casa Editrice Ambrosiana. Structure determination of organic compounds, practical exercises E. Rossi, D. Nava, G. Abbiati, G. Celentano, S. Pandini. Casa Editrice Edises
A. Randazzo. Guida pratica all'interpretazione di spettri NMR. Casa Editrice Loghia
A. Randazzo. Guida pratica all'interpretazione di spettri NMR. Casa Editrice Loghia
Analytical and pharmacodynamic characterization of biopharmaceuticals
Course syllabus
Biopharmaceuticals and in particular therapeutic proteins produced using recombinant DNA technology are complex, heterogeneous molecules, and subject to a variety of enzymatic and / or chemical modifications that can occur during expression, purification, and long-term storage.
The course consists of two parts. The first part is concerning the structural characterization of biopharmaceuticals, whereas the second part is concerning their pharmacodynamic characterization.
The structural characterization of latest generation biopharmaceuticals will be taught using as educational example the characterization of antibody-drug conjugates (ADCs) as a comprehensive educational path to teach the most popular analytical techniques and methodological approaches. The program will include the fundamentals of:
1) Chromatography (high performance liquid chromatography)
2) Electrophoresis (gel and capillary electrophoresis)
3) UV-Vis spectroscopy (absorption and emission techniques)
4) Mass spectrometry (ESI and MALDI ionization)
The fundamentals of hyphenation of such analytical techniques and the principles of sample preparation and purification necessary for sample analysis will be discussed as well.
Concerning the pharmacodynamic characterization of biopharmaceuticals, determination of the efficacy of such products refers to the quantitative measurement of their biological activity. Biological activity is, in turn, a critical quality attribute; therefore, efficacy assays are an essential component of the quality control of the entire production process ("the product is the process"). During the course, different biological activity testing procedures will be illustrated, including:
1) Biochemical tests (e.g., enzymatic and immunoenzymatic assays)
2) Assays based on the use of cell cultures
3) Assays based on the use of experimental animals (traditional and conditional transgenic models, etc.)
Furthermore, information will be provided on the set up of new assays based on the use of human iPS (induced pluripotent stem cells), organoids, gene editing and CAR-T technologies, as well as on the National Centers for drug quality control, and on the local and national Organisms responsible for the regulation of animal experimentation.
The course consists of two parts. The first part is concerning the structural characterization of biopharmaceuticals, whereas the second part is concerning their pharmacodynamic characterization.
The structural characterization of latest generation biopharmaceuticals will be taught using as educational example the characterization of antibody-drug conjugates (ADCs) as a comprehensive educational path to teach the most popular analytical techniques and methodological approaches. The program will include the fundamentals of:
1) Chromatography (high performance liquid chromatography)
2) Electrophoresis (gel and capillary electrophoresis)
3) UV-Vis spectroscopy (absorption and emission techniques)
4) Mass spectrometry (ESI and MALDI ionization)
The fundamentals of hyphenation of such analytical techniques and the principles of sample preparation and purification necessary for sample analysis will be discussed as well.
Concerning the pharmacodynamic characterization of biopharmaceuticals, determination of the efficacy of such products refers to the quantitative measurement of their biological activity. Biological activity is, in turn, a critical quality attribute; therefore, efficacy assays are an essential component of the quality control of the entire production process ("the product is the process"). During the course, different biological activity testing procedures will be illustrated, including:
1) Biochemical tests (e.g., enzymatic and immunoenzymatic assays)
2) Assays based on the use of cell cultures
3) Assays based on the use of experimental animals (traditional and conditional transgenic models, etc.)
Furthermore, information will be provided on the set up of new assays based on the use of human iPS (induced pluripotent stem cells), organoids, gene editing and CAR-T technologies, as well as on the National Centers for drug quality control, and on the local and national Organisms responsible for the regulation of animal experimentation.
Teaching methods
The course is delivered by classroom lectures, in which the topics are illustrated both with by slides and by exercises on the blackboard. Attendance to classes is highly advised. To promote student's engagement some topics are delivered through flipped classrooms, breakout rooms, as well as interactive live quizzes and Q&A sessions.
Teaching Resources
Slides of the room lessons and other material available online at the Ariel website of the course
Antibody-Drug Conjugates, Methods and Protocols, edited by L.NathanTumey (2020), ebook available at the university online library service, ISBN: 1-4939-9929-X; 1-4939-9928-1.
Analytical Characterization of Biotherapeutics (capitoli 1,2 e 7), edited by Jennie R. Lill and Wendy Sandoval (2017), ebook available at the university online library service, ISBN: 1-119-38442-7; 1-119-38443-5; 1-119-05310-2.
Antibody-Drug Conjugates, Methods and Protocols, edited by L.NathanTumey (2020), ebook available at the university online library service, ISBN: 1-4939-9929-X; 1-4939-9928-1.
Analytical Characterization of Biotherapeutics (capitoli 1,2 e 7), edited by Jennie R. Lill and Wendy Sandoval (2017), ebook available at the university online library service, ISBN: 1-119-38442-7; 1-119-38443-5; 1-119-05310-2.
Analytical and pharmacodynamic characterization of biopharmaceuticals
BIO/14 - PHARMACOLOGY
CHIM/08 - PHARMACEUTICAL CHEMISTRY
CHIM/08 - PHARMACEUTICAL CHEMISTRY
Lessons: 48 hours
Professors:
Abbracchio Maria Pia, Regazzoni Luca Giovanni
Pectroscopic methods in bioorganic chemistry
CHIM/06 - ORGANIC CHEMISTRY - University credits: 3
Lessons: 24 hours
Professor:
Pini Elena Renata Elvira
Educational website(s)
Professor(s)
Reception:
By appointment
Via Golgi 19. Entrance C, IV floor
Reception:
on demand, to be scheduled via email
Via L. Mangiagalli 25, first floor, room 1044