PEGylated proteins quantification strategies by LC-MS/MS

Quantification by LC-MS/MS of the PEGylated protein Pegvisomant in the ng/mL range without prior protein purification or enrichment   

Quantification of PEGylated proteins in plasma by LC–MS/MS encompasses challenges that may not be encountered in quantification of non-PEGylated proteins and requires careful consideration. Depending on the method of conjugation, PEGylation at targeted amino acids may not be stoichiometric or consistent, therefore requiring a surrogate peptide that is void of potential PEGylation sites. This selection process reduces the pool of candidate signature peptides that are amenable for quantification.

What will you learn?

  1. How to address Bioanalytical challenges with PEGylated proteins.
  2. Choosing the right surrogate peptide in PEGylated proteins.
  3. Reduce your sample preparation expenses by optimizing of sample preparation steps for PEGylated proteins.
  4. Optimal tuning of LC and MS parameters to overcome the challenges of PEGylated protein quantification.
  5. Case study: development of rugged and efficient bioanalytical methods for a PEGylated protein.

Who should attend?

Scientists transitioning from bioanalysis of small molecules to large molecules

Panelist’s biography

ABSciex_March_author

 

 

 

Dr. Fabio Garofolo
Vice President of Bioanalytical Services
Algorithme Pharma, Laval, Quebec, Canada

Dr. Fabio Garofolo has been working in the pharmaceutical and analytical fields for more than 22 years. Capitalizing on an extensive occupational background within his specialization field, he has also been involved to help scientific non-profit organizations (AAPS, ASMS, CFABS, CVG, GBC, GCC and NBC) with the mission to promote discussion on the pharmaceutical practices.

Dr. Garofolo has progressed throughout in his career in both the pharma/biotech and CRO industries and benefited from successfully overcoming the prevalent challenges in drug development. Since 2005 he has held the position of Vice-President Bioanalytical Services at Algorithme Pharma. Dr. Garofolo has over 130 publications & presentations in international conferences. He developed around 300 innovative analytical methods. He designed and invented 3 innovative bioanalytical approaches and is the recipient of the following Lilly awards: Achievement (2001); Global (2002); Emmerson (2003). His current interests include analysis of large molecules by LCMS, HRMS, DBS, and Emerging Technologies.