Posters

Intact 20kDa Extracellular Domain of APO2L/TRAIL Bioanalysis by HRMS: A Potential Cancer Therapeutic Protein

Jean-Nicholas Mess1 and Fabio Garofolo1

1Algorithme Pharma Inc., Laval, Québec, CANADA

NOVEL ASPECT:

Recombinant APO2L/TRAIL, typically analyzed by LBA, was successfully quantified as intact protein by High Resolution Mass Spectrometry (HRMS).

INTRODUCTION:

APO2L/TRAIL (Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand) is the first recombinant human protein that selectively induces apoptosis or programmed cell death in cancer cells while sparing normal cells. Although typically quantified by ligand binding assays (LBA), high resolution mass spectrometry (HRMS) instruments offer an alternative for the quantification of large molecules to support pharmacokinetic study in early drug discovery. Indeed, HRMS assays are generally faster to develop since they do not require specific reagents such as highly selective monoclonal antibodies. Moreover, due to high resolution and extended mass range of QTOF, intact protein quantification may be considered as a valuable option. APO2L/TRAIL was analyzed using intact quantification without tryptic digestion by HRMS.

Abstract Number/
Poster Board number:

294 Poster Board -506

Presenting Author:
Fabio Garofolo

Session Title:
Cell Death/Apoptosis

Date & Time:
March 24, 2014
9:30 AM to 12:30 PM

Location:
CC Exhibit Hall
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Bioanalysis of Exenatide: Intact Versus Signature Peptide Approach to Reach Optimal Sensitivity in Large Molecule Quantification by LC-MS

Jean-Nicholas Mess1 and Fabio Garofolo1

1Algorithme Pharma Inc., Laval, Québec, CANADA

NOVEL ASPECT:

Direct comparison of two different quantification approaches; intact versus signature peptide, for the quantification of Exenatide in human plasma.

INTRODUCTION:

LC-MS is an emerging technique for the quantification of large molecules like therapeutic peptides, proteins and biomarkers. However, due to the limited mass range of triple quadrupole mass spectrometers, these analytes are typically monitored as multicharged species. This dispersion of MS signal combined with poor fragmentation (CID) efficiency may affect the achievable LOQs. Another approach is to generate smaller signature peptides through tryptic digestion. These tryptic peptides will generally show less charge state distribution and clearer CID pattern than the intact analyte. However, the efficiency of the digestion process could be poor and compromise its sensitivity. In this study, both approaches will be compared for the quantification of exenatide in human plasma.

Abstract Number/
Poster Board number:

879 Poster Board -248

Presenting Author:
David Leroux-Petersen

Session Title:
Pharmacokinetics and Disposition

Date & Time:
March 25, 2014
9:00 AM to 12:30 PM

Location:
CC Exhibit Hall
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Glucagon Bioanalysis by LC-MS: “Unprecedented Level of Sensitivity (10pg/mL) for a Novel Formulation”

Jean-Nicholas Mess1, Louis-Philippe Morin1, Mauro Aiello2, Xavier Misonne2, Gary Impey2, Johnny Cardenas2, Josee Michon1, and Fabio Garofolo1

1Algorithme Pharma Inc., Laval, Québec, CANADA
2AB SCIEX, Concord, Ontario, CANADA

NOVEL ASPECT:

Development of a LC-MS/MS method is more sensitive than LBA for challenging quantification of Glucagon at low pg/mL level in new formulation.

INTRODUCTION:

Ligand Binding Assay (LBA) is currently the most common approach for Large Molecule quantification. However, LBA may suffer from cross reactivity and lack of specificity and selectivity for very low quantitation analysis. In the past several years, LC-MS based assays showed to be as effective and also complementary to LBA for Large Molecule quantification. Currently, we are working on a study for a novel formulation of glucagon which should be easier to administer than the currently available formulations resulting in a higher standard of Type 1 diabetes patient care. The present work describes the challenges and solutions encountered using novel mass spectrometers to reach unprecedented level of sensitivity for the analysis of low plasma concentration of glucagon.

Abstract Number/
Poster Board number:

880 Poster Board -249

Presenting Author:
Marie-Hélène Raigneau

Session Title:
Pharmacokinetics and Disposition

Date & Time:
March 25, 2014
9:00 AM to 12:30 PM

Location:
CC Exhibit Hall
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Advanced Use of High Resolution Mass Spectrometry (HRMS) to Overcome Triple Quadrupole Limitations in Large Molecules Quantification

Louis-Philippe Morin1, Jean-Nicholas Mess1, Milton Furtado1 and Fabio Garofolo1

1Algorithme Pharma Inc., Laval, Québec, CANADA

NOVEL ASPECT:

The use of HRMS (QTOF) to overcome the limitation of triple quadrupole for development of large molecule bioanalytical assay.

INTRODUCTION:

The quantitation of large molecules by LC-MS has gained great popularity in the bioanalytical industry. Although triple quadrupoles are sensitive and reliable instruments having advantages over standard immunoassays (LBA), it is still not perfectly adapted to large molecule quantification. Since quadrupole instruments have limitations such as low resolution and limited mass range, the use of the new generation of HRMS such QTOF and Orbitraps is becoming an accepted alternative. In fact, HRMS are much more adapted to resolve some issues associated with large molecule quantitation. This research discusses two Large Molecule case studies (somatostain and enfurvirtide) where the use of a HRMS (TripleTOFTM5600) was found more sensitive than a triple quadrupole (QTRAP®5500).

Abstract Number/
Poster Board number:

881 Poster Board -250

Presenting Author:
Robert Sabelli

Session Title:
Pharmacokinetics and Disposition

Date & Time:
March 25, 2014
9:00 AM to 12:30 PM

Location:
CC Exhibit Hall
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