Bioanalysis Webinar: LC and LC-MS of Intact Protein Biotherapeutics and their Variants and Panel Discussion
1:00 p.m. – Dr. Lijuan Kang, Sr. Scientist, Janssen Pharmaceutical Companies of Johnson & Johnson: Current Chromatography Developments in Intact Protein (Top-Down) Therapeutics Bioanalysis by LC-HRMS
1:30 p.m. – Dr. Barry Boyes, Vice President of R&D, Advanced Materials Technology, Inc.: A Useful Reversed-Phase LC Method Development Strategy for Large Protein Variants
2:00 p.m. – Open Panel Discussion and Q&A Panelists: • Dr. Naidong Weng, Scientific Director, Janssen Fellow, and Head of East Coast Bioanalytical Chemistry and Pharmacokinetics, Janssen Research & Development • Dr. Tao Jiang, R&D Manager, Mallinckrodt Pharmaceuticals • Dr. Thomas Waeghe, Senior Scientist, MAC-MOD Analytical • Dr. Barry Boyes, Vice President of R&D, Advanced Materials Technology, Inc. • Dr. Michael Dong, Principal, MWD Consulting
Registration link: https://register.gotowebinar.com/register/9099462980534316302?source=Email
|Abstract: Current Chromatography Developments in Intact Protein (Top-Down) Therapeutics Bioanalysis by LC-HRMS|
|Intact (top-down) protein bioanalysis is being developed as a complementary approach to bottom-up analysis for protein therapeutics. It has unique advantages in providing simultaneous catabolite identification and quantitative bioanalysis. However, this approach also faces challenges such as poor chromatographic peaks and inadequate assay sensitivity. In current work, different conditions such as HPLC columns were screened to optimize chromatography for the intact protein analysis. To improve the assay sensitivity, micro-flow LC-MS has been explored. Micro-flow LC-MS is known to enhance sampling and ionization efficiency and has been demonstrated to improve sensitivity for small molecules and peptides, but so far has not been reported for the intact protein bioanalysis. This talk will discuss important considerations when applying micro-flow LC for intact protein bioanalysis and compare preliminary results obtained under different configurations. The data showed that application of micro-flow LC-MS, while being successfully used for the bottom-up protein bioanalysis, warranted further exploration since many parameters such as column chemistry and system configuration affect the performance of micro-flow LC-MS.|
|Abstract: A Useful Reversed-Phase LC Method Development Strategy for Large Protein Variants|
The number of biotherapeutic drugs including monoclonal antibodies (mAbs) and their related products, such as antibody-drug conjugates (ADCs), have been growing rapidly, particularly in the last decade. The development and commercialization of these complex entities is a complicated process and requires a significant amount of effort and capability — especially in chromatographic separations. This has created a need for chromatographers to have effective strategies for LC separations of large (>50,000 MW) and complex proteins, such as mAbs and their variants. In this webinar, we will discuss how to approach your method development strategy by adjusting various parameters of your separation to ensure that you are seeing more of the variants of your complex proteins. These parameters include pore size, bonded phase chemistry, mobile phase modifiers, mobile phase additives, and column temperature. Systematic selection and adjustment of these variables leads to improved resolution of highly similar protein variants. An example of this approach is high-resolution intact protein LC–MS analysis of difficult IgG mixtures, such as the complex mixtures of lower abundance free sulfhydryl variants present in therapeutic mAbs.
Please register for Applications of Empore membrane SPE techniques in LC mobile phase cleanup and the sample preparation for Proteomics
When: May 6, 2020 12:00 PM EDT at:
After registering, you will receive a confirmation email containing information about joining the webinar.
Guotao Lu, MS Chemistry
Empore Product Manager
CDS Analytical, LLC.
In this seminar, we explore the power of Empore membrane SPE technique for two unique applications in chromatography. The first application is to use Empore Disks to clean up LC mobile phase to improve LC method development efficiency by eliminating the ghost peaks and interferences. The second application is Empore Disks as StageTips in Proteomics.
Since its first invention by Professor Mann’s group from Germany in 2003, Empore StageTips technique has been widely used in Proteomics research. The main functions of Empore StageTips are protein desalting and peptide purification. Google Scholars showed over 6,000 publications about StageTips application in Proteomics. Researchers have used Empore StageTips for routine proteomics sample preparation, as well as special applications in phosphous-proteomics, single-cell proteomics and global proteomics. The unique advantages of Empore C18 StageTips for high-pH fractionation in proteomics are particularly addressed in this seminar.
Key Learning Objectives:
- Utilize Empore SPE Disks to clean-up mobile phase to eliminate interferences in LC method development
- Empore SPE Disks to make StageTips for protein desalting and peptide purification in proteomics
- Advantages of Empore C18 StageTips for high-pH fractionation in proteomics
Who Should Attend:
- Method developers for HPLC methods in pharmaceutical, chemical, clinical, environmental, agrichemical, contract research organization, university and governmental laboratories.
- Proteomics researchers
About CDS Analytical:
CDS Analytical has celebrated its 50th anniversary. We have been offering various GC sample introduction instrument, including 6150, 6200, 5200HPR Pyroprobe for Pyrolysis, 9300 series and 7550S for Thermal Desorption, 7000E and 7000C for Purge & Trap and Dynamic Headspace. CDS Analytical acquired 3M EmporeTM in 2019 as a new product line in Solid Phase Extraction.