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The CHROMacademy Essential Guide Webcast:
Analysis of Next Generation Protein Biopharmaceuticals

Thursday 17th November 2016,
8:00am PDT/ 11:00am EDT/ 4:00pm BST/ 5:00pm CEST

 
 

This webcast will focus on next generation biopharmaceuticals, for example, biosimilars, biobetters, and antibody drug conjugates.  We will look at how chromatographic and mass spectrometric techniques can be used to distinguish differences between originator and next generation biopharmaceuticals and which properties are assigned as Critical Quality Attributes (CQAs).

Presented by

Dr. Andrew Coffey (Snr Applications Chemist, Agilent Technologies)

Dr. Dawn Watson (CHROMacademy Technical Expert, Crawford Scientific)

Topics Covered

  • What are next generation protein biopharmaceuticals?
  • Biosimilars - How similar is similar enough?
  • Biosimilars versus biobetters - distinguishing similar from better
  • Analytical tools for assessing similarity
  • Techniques for analyzing bispecific antibody drug conjugates
  • MS based techniques for the comparison of originator, biosimilar, and biobetter antibodies

 

Presenter Information »

 
 

Analysis of Next Generation Protein Biopharmaceuticals

This month’s webcast focused on the analysis of next generation protein biopharmaceuticals.  The CHROMacademy material provided in this month’s tutorial will help you understand the analytical tools which are employed in the analysis of protein biopharmaceuticals and what information each technique provides.

Our Quick guides, CHROMacademy course content, and archived webcasts and tutorials provide practical information on implementing a variety of analytical techniques to the analysis of protein biopharmaceuticals, including chromatographic conditions, how to select the correct column, and the effect method parameters can have on data.

 

eLearning Modules


 
Biopharmaceuticals

An introduction to the biopharmaceutical market place and types of biopharmaceuticals.

Generic Biopharmaceuticals | Glycoengineering | Bispecific Antibodies | Antibody Drug Conjugates

 
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 Post-Translational Modifications (PTMs) | Deamidation
Oxidation | Peptide Mapping in a Routine Environment

 
HILIC

This module provides information on how hydrophilic interaction liquid chromatography is used to analyze protein biopharmaceuticals, in particular for the analysis of glycans.  Typical glycan mapping conditions will be provided as well as details on what information can be gained from this technique.

Introduction to HILIC for Biopharmaceuticals | Typical Glycan Mapping HILIC Conditions
Glycan Analysis During Biosimilar Development

 
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 »

 
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 »

 
Advances in Full Characterization of Protein Biopharmaceutical Post Translational Modifications

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.

Advances in Full Characterization of Protein Biopharmaceutical Post Translational Modifications »

 
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 »

 
 
 
 
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Andrew Coffey, PhD;
Snr Applications Chemist,
Agilent Technologies

Andrew joined Polymer Laboratories in 1989 after completing his PhD in peptide synthesis at Wolverhampton, UK.

He progressed from Senior Scientist to Product Manager looking after the HPLC column product line before focusing on Solid Phase specialty particle businesses.

Adding Biosolutions and Flash chromatography consumables product lines to his portfolio during the acquisition by Varian and then Agilent Technologies, he joined the Biocolumns Applications Team in August 2011.

Andrew has considerable experience in reversed phase, ion-exchange and size exclusion liquid chromatography method development, technical support and troubleshooting and has a “hands on” attitude to problem solving!


Dr. Dawn Watson, CHROMacademy Technical Expert,
Crawford Scientific

Dawn received her PhD in synthetic inorganic chemistry from the University of Strathclyde, Glasgow.  The focus of her PhD thesis was the synthesis and application of soft scorpionate ligands.  As well as synthetic skills, this work relied on the use of a wide variety of analytical techniques, such as, NMR, mass spectrometry (MS), Raman spectroscopy, infrared spectroscopy (IR), UV-visible spectroscopy, electrochemistry, and thermogravimetric analysis. 

Following her PhD she spent two years as a postdoctoral research fellow at Princeton University studying the reaction kinetics of small molecule oxidation by catalysts based on Cytochrome P450.  In order to monitor these reactions stopped-flow kinetics, NMR, HPLC, GC-MS, and LC-MS techniques were utilized.  Prior to joining the Crawford Scientific and CHROMacademy technical team she worked for Gilson providing sales and support for the entire product range including, HPLC (both analytical and preparative), solid phase extraction, automated liquid handling, mass spec, pipettes, and laboratory consumables.