No thanks! I would like to know more about CHROMacademy

 Over 3000 E-Learning topics / 5000 Articles & Applications
 

The CHROMacademy Essential Guide Webcast:
Advances in Full Characterization of Protein Biopharmaceutical Post Translational Modifications.

Thursday 22nd September 2016,
8:00am PDT/ 11:00am EDT/ 4:00pm BST/ 5:00pm CEST

 

Post translational modification (PTMs) which are most characteristic in protein biopharmaceuticals include glycosylation (which comprises galactosylation, fucosylation, high mannose derivatives and sialylation), oxidation, phosphorylation, sulfation, lipidation, disulfide bond formation and deamidation, as well as (to a more limited extent) carboxylation, hydroxylation, and amidation; with many biopharmaceuticals having a combination of two or more PTMs.  PTMs need to be monitored during numerous stages of the manufacturing process and require identification, control of their levels, and assessment of their impact on the protein.  In order to do this many chromatographic (reversed phase and HILIC HPLC) and mass spectrometric (MS) techniques are employed.  This webcast will consider the types of modifications which can be assessed and what methods should be used to identify them, how various MS scanning techniques can be used for PTM analysis, as well as considering if the analysis mode itself can induce post translation modifications (collision induced dissociation vs. electron transfer dissociation (ETD) and electron capture dissociation (ECD)).  Finally, the use of LC-MS for protein determination of antibody-drug conjugates in biological matrices will be discussed.

Presented by

Dr. Donna Potts, Biopharma Market Manager – EMEA, Agilent Technologies

Tony Taylor, Technical Director, Crawford Scientific

Topics Covered

  • What post translational modifications can be assessed?
  • What effect do post translation modifications have on protein biopharmaceuticals?
  • Reversed phase HPLC analysis of PTMs
  • Utilizing different MS scanning functions to enable post translational modification events
  • How the analysis method can alter the post translation modification - alternatives to classical collision induced dissociation (top down analysis)
  • Host cell protein analysis using LC-MS
  • Protein quantification using LC-MS (pharmacokinetic studies) - protein determination, in particular ADC determination, in biological matrices

 

Presenter Information »

 
 

Advances in Full Characterization of Protein Biopharmaceutical Post Translational Modifications

The CHROMacademy material provided in this month’s tutorial will help you understand which post translational modifications need to be considered in relation to biopharmaceuticals and the techniques that can be used for their analysis.

Our Quick guides, CHROMacademy course content, and archived webcasts and tutorials will provide you with the knowledge you require to gain an understanding of the post translational modifications which occur in biopharmaceuticals, the effect they have on their function, and which chromatographic techniques can be used to characterize them.  Practical starting chromatographic conditions will be discussed, as well as a closer look at specific parameters which can impact separations.  Furthermore, you will be introduced to other techniques for biopharmaceutical analysis, including ion exchange and size exclusion chromatography.

Watch the webcast

eLearning Modules


 
Reversed Phase - Introduction and Peptide Level Analysis

Everything you need to know about using reversed phase analysis for characterizing peptide fragments of biopharmaceuticals, including mAb digestion using trypsin, what a typical peptide may looks like, and which post-translational modifications can be identified and how.

Analysis at the Peptide Level | Common mAb PTMs | Glycoprofile

 
Reversed Phase - Peptide Mapping LC Conditions and Effects

This module will provide you with practical details of how to implement peptide mapping of biopharmaceuticals, from initial conditions to considering the effect of reversed phase parameters such as mobile phase additives, temperature, gradient slope, and column dimensions on the separation.

Typical Peptide Mapping Conditions | Mobile Phase Additives | Gradient Slope/Time

 
mAbs - Introduction to Monoclonal Antibodies

Learn about the fundamentals of monoclonal antibodies (mAbs), such as, how they are synthesized in vivo, key structural features, and the significance of post-translational modifications in relation to  mAb function.

Introduction to Monoclonal Antibodies | mAb post-translational modifications
Disulfide Bond Scrambling, Isoforms, and Aggregate Formation | Glycoform Variation in N-Linked Glycans

 
Introduction to Biopharmaceuticals

Biopharmaceuticals come in a range of formats, including, monoclonal antibodies (mAbs), bispecific antibodies, and antibody-drug conjugates (ADC).  This module introduces the biopharmaceutical market place and provides information on the different forms of biopharmaceuticals and how they work.

Antibody Drug Conjugates

 
Quick Guides

Reversed Phase HPLC for the Analysis of Biomolecules

Commonly reversed phase (RP), hydrophilic interaction (HILIC), ion exchange (IEX), and size exclusion (SEC) chromatographic modes will be combined to allow full characterization of complex biomolecules. The following article will introduce the fundamentals of biopharmaceutical analysis and cover the use of reversed phase HPLC in their analysis.

Reversed Phase HPLC for the Analysis of Biomolecules »

 
Do you know your peptides from your proteins?

A little bit of fun to test your knowledge of biopharmaceuticals and their analysis.

Biopharmaceuticals Quiz »

 
HILIC, IEX, and SEC for the Analysis of Biomolecules

The enhanced complexity and variability that comes from the size of biopharmaceuticals, allied with the intricacy of the production process, mean chromatography is employed to a much greater extent during production and release testing. This article will examine the use of HILIC, IEX, and SEC for their full characterization.

HILIC, IEX, and SEC for the Analysis of Biomolecules »

 
Selecting Columns and Initial Conditions for IEX and SEC

Biopharmaceutical compounds are genetically engineered from living cells and have molecular weights in the range 2,000-2,000,000 Da with 10-2,000 reactive groups and are often comprised of a mixture of closely related variants; therefore, there is a wider degree of variability (a degree of heterogeneity) in comparison with small molecule pharmaceuticals.

Selecting Columns and Initial Conditions for IEX and SEC »

 
The Analysis of Post Translational Modifications Using LC-MS/MS

This webcast will explain the principles behind the analysis of a number of key post-translational modifications (PTMs) using LC-MS/MS techniques. The use of bottom-up proteomics and different MS scanning techniques to characterize PTMs such as, phosphorylation, acetylation, ubiquitination and SUMOylation will be covered.

The Analysis of Post Translational Modifications Using LC-MS/MS »

 
Characterization of Protein Biopharmaceuticals

Protein biopharmaceuticals, such as monoclonal antibodies and recombinant proteins, are currently in widespread use for treatment of various life-threatening diseases including cancer, autoimmune diseases, diabetes, anemia, etc. Protein therapeutics have a complexity far exceeding that of small molecule drugs, hence, unraveling this complexity represents an analytical challenge. This webcast will highlight the power of liquid chromatography combined with mass spectrometry in the characterization of protein biopharmaceuticals.

Characterization of Protein Biopharmaceuticals »

 
HPLC Techniques in Biopharmaceutical Analysis

The enhanced complexity and variability that comes from the size of biopharmaceuticals, allied with the intricacy of the production process, mean chromatography is employed to a much greater extent during production and release testing. This Essential Guide webcast will provide details of why differing modes of chromatography are required in the research, preparation, and analysis of biopharmaceuticals and when they would be typically used.

HPLC Techniques in Biopharmaceutical Analysis »

 
 
 
loading data
loading data
loading data
loading data
loading data
Home | About UsContact Us | SubscribeTerms and Conditions | Advertise | Privacy Policy |

loading data

loading data

loading data

 

loading data


loading data

 

Tony Taylor
Technical Director, Crawford Scientific

Tony comes from a pharmaceutical background and has many years’ research and development experience in small molecule analysis and bio-analysis using LC, GC and hyphenated MS techniques.

Tony is actively involved in method development within the analytical services laboratory at Hall Analytical – which supplies contract research in Chromatography and Mass Spectrometry. He continues to research in novel separation technologies, chromatographic method development and structural characterisation, especially in the areas of Extractable and Leachable analysis and Bio-Chromatography.

Tony is the Technical Director of the CHROMacademy and has spent the past 17 years as a trainer and developing on-line education materials in analytical chemistry techniques.


Dr. Donna Potts,
Biopharma Market Manager
EMEA,
Agilent Technologies

As the European Biopharma Market Manager for Agilent Technologies, Donna Potts is responsible for business development for the Agilent Biopharma portfolio, including high performance LC, LC-MS, bio-columns, and automation systems, alongside associated applications. Using her broad knowledge in this market-space, Donna works closely with Biopharma customers to identify and develop appropriate analytical solutions to help solve their workflow limitations, enabling improved results and productivity.

Donna has worked extensively with biomolecules during her career, with a strong emphasis on analysis of proteins, antibodies and glycans using LC-MS. She received her MChem (Hons) and Doctorate degree from UMIST, UK with a thesis on protein quantitation using mass spectrometry, followed by post-doctorate study at Boston University School of Medicine in the group of Professor Catherine Costello. Since leaving academia, Donna has worked as an application specialist for major instrument vendors, focusing on Life Science applications including proteomics and Biopharma.