Protein Therapeutic Drugs
Analysis and Characterization

Instrumental Tools, Techniques and Tips for the Characterization and Analysis of Biotechnology-derived Polypeptide Therapeutics

מרצה: Mr. David Carr

תאריך: יפורסם בהמשך
מיקום הקורס: כפר המכביה, רמת גן
מידע כללי
משך הפעילות: 4 מפגשים, יום שלם
שעות: 08:30-17:00
עלות: 6,450 ש"ח + מע"מ
נקודות זכות: 3
מק''ט: 17825
שפת הקורס: אנגלית
על הקורס

New regulations issued by the FDA on May 14, 1996 substantially changed the way in which biotechnology-derived pharmaceuticals are regulated, placing greater emphasis on the methodologies used to analyze biotechnology-derived polypeptide therapeutics. Fundamentals in Analytical Biotechnology immerses those attending in the practical aspects of characterizing and analyzing therapeutic proteins and presents in an adsorbing manner the fundamental principles and practical uses of these three analytical technologies. Topics extraneous to protein analysis are eliminated, allowing the course to focus on the points of each technology essential in protein characterization. The materials can become almost overwhelming because of the sheer weight of information presented, and yet all points become clear because of the structure of the class and the ability of the instructor to present each topic clearly.

Those attending leave with a clear sense of where each technology is used and how to most effectively implement each technology in developing and using effective, efficient analytical methods for characterizing and analyzing therapeutic proteins.

The Analysis and Characterization of Protein Therapeutics Drugs is a condensed and focused course of instruction on the instrumental analysis and characterization of polypeptides, emphasizing three important instrumental analytical technologies as they relate to the analysis and characterization of polypeptides.

  • Protein Structure and Function. Modifications to proteins that may affect function or potency.
  • Chromatography, including reversed-phase HPLC, ion exchange and other techniques related to protein analysis
  • Capillary Electrophoresis, a technique used for the analysis of charge variants, antibodies and glycosylation
  • Mass Spectrometry, a powerful technique for the analysis and characterization of protein modifications.
קהל היעד
  • Young scientists starting to work with protein pharmaceuticals
  • Experienced pharmaceutical analysts who are starting to work with protein drugs
  • Regulatory scientists working with protein pharmaceuticals
  • Research and Development scientists developing biopharmaceuticals
Protein Therapeutic Drugs
Analysis and Characterization
התועלת שתופק

This is a three and a half day classroom lecture course focusing on the structure and function of proteins, modifications to proteins that may affect their function, and the fundamental principles and practical use of three major instrumental techniques – Chromatography, especially reversed-phase HPLC (other chromatography techniques are also discussed), capillary electrophoresis and mass spectrometry - in the analysis and characterization of biotechnology-derived polypeptide biopharmaceuticals.

השאר פרטים למידע נוסף
  • שם מלא
  • דוא''ל
  • טלפון
  • הערות
  • שלח
תכני הקורס

Day 1
Introduction to the course including a discussion of the history of the regulation of protein therapeutics.
Protein Structure and Function

  • Amino acids. Structure and character.
  • The peptide bond
  • The nature of the side-chains of the common amino acids.
  • The interactions which form and stabilize polypeptide
  • Secondary structure. The alpha-helix and beta-sheets
  • Protein tertiary structure and its role in biological activity
  • How primary and secondary structure lead to tertiary structure.
  • Protein quaternary structure
  • Why structure matters

Protein Degradations and Post-translational Modifications

  • Post-translational modifications such as glycosylation, the addition of oligosaccharides to polypeptides
  • Deamidation and oxidation and their effects on protein function and biological activity, protein stability
  • Changes to tertiary structure, Denaturation and aggregation.
  • Engineered proteins, Protein conjugates, Monoclonal antibody conjugates Pegylation

Day 2
Reversed-phase High Performance Liquid Chromatography

  • Basic theory and mechanism of polypeptide separations
  • Developing an analytical method for the analysis of polypeptides. Deciding on the right column. Selecting the elution solvent including the organic modifier, the ion-pairing reagent, the pH, the flow rate and the gradient conditions. Detection mode. The affect of Temperature
  • Developing a reversed-phase HPLC assay. The steps in developing a robust analytical method
  • Tips for the effective use of reversed-phase HPLC in polypeptide separations
  • Examples of how reversed-phase HPLC is used to characterize and analyze protein degradations, post-translational modifications and other important protein attributes

Other types of chromatography in polypeptide analysis

  • Ion Exchange Chromatography (Charge variants)
  • Hydrophobic interaction chromatography (Protein modifications)
  • Size exclusion chromatography (Aggregation and Pegylation)
  • Chromatography used in the analysis of glycosylation: High pH Anion Exchange, Normal phase and HILIC

Day 3
Capillary Electrophoresis

  • Basic principles of CE. What is CE? Comparison with HPLC and gel electrophoresis. Understanding basic terms in CE: Mobility, Migration, Separation efficiency, and Electroosmotic flow. Basic CE instrumentation. Sample detection. Sample injection. The capillary and temperature control
  • Capillary Zone Electrophoresis. What is capillary zone electrophoresis? Developing a CZE method including selecting the capillary, choosing the best buffers, the importance of pH, the role of the sample matrix, the use of buffer additives and choosing the correct voltage. Sample stacking, what it is and how to use it
  • Capillary Isoelectric focusing. A description of Capillary Isoelectric focusing. The steps in CIEF. Practical use of CIEF
  • Capillary Replacable Gel Electrophoresis (CE-SDS). A brief discussion of CE-SDSand how it is used in bioanalytical separations
  • How capillary electrophoresis is used to meet the needs and challenges of polypeptide characterization and analysis

Mass Spectrometry

  • Basic principles and terminology of mass spectrometry
  • Ionization sources: Matrix-Assisted Laser Desorption Ionization (MALDI) Electrospray. For each ionization source, a discussion of selecting conditions, the affect of concomitant species and optimizing performance.
  • Mass analyzers: Quadrupole. Ion Trap. Time-of-Flight. Orbitrap
  • Interfacing mass spectrometry to HPLC and CE

Day 4
Mass spectrometry continued

  • Fragmentation of ions in mass spectrometry.
    Collisionally-Induced Dissociation (CID). In-source CID in electrospray.
  • Applications.
    How mass spectrometry is used to meet the needs and challenges in the characterization and analysis of protein therapeutic drugs. Determination of molecular weight of proteins by electrospray-MS. The role of mass spectrometry in peptide maps. Sequencing peptides by electrospray CID MS and by MALDI-MS. How mass spectrometry is used to determine protein modifications and degradations.
  • The role of mass spectrometry in glycan analysis.
אודות המרצה

Mr. David Carr

David Carr is a graduate of the University of California, Berkeley and did graduate work at the University of California, San Diego, in biochemistry and molecular biology. He began working in the field of High Performance Liquid Chromatography (HPLC) in 1971 at Varian Associates, a pioneer in this field.
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