Abstract 2064: Mass Spectrometry Imaging of therapeutic antibodies: Distribution of unlabeled trastuzumab in CB.17 SCID mice implanted with the human breast BT474 xenograft
Authors: Bonnel David , Hall Chassidy , Mullin Robert J. , Simon Kathryn A. , Stauber Jonathan 
Published: May 2015

Introduction

Mass Spectrometry Imaging (MSI) is an emerging technique in preclinical analysis that allows for analysis of the distribution and the quantification of target molecules in tissue sections. The main advantage of this imaging technology is the detection of the molecule of interest in tissues without labelling and with high specificity. MSI is now routinely used for small molecule distribution analysis in lead optimization process, and in PK/PD or toxicity studies. Due to the mass range limitation of the instrument analyzer one of the main challenges is the detection of administered large molecules such as therapeutic antibodies. . In this study, we describe the detection and the distribution of trastuzumab, a monoclonal antibody that recognizes the HER2/neu receptor, in a BT474 human breast xenograft model.

Experimental procedures

BT474 tumor fragments are implanted in female CB.17 SCID mice. When the tumors reach 80-120 mm3, the trastuzumab treatment of 20mg/kg with eight bi-weekly intraperitoneal injections was initiated. The endpoint was fixed at day 30 post first administration. At 50 hours post 1st, 5th and 8th doses vehicle and trastuzumab treated animals were sacrificed and tumor, plasma and carcass were collected.

One section of each tumor tissue was first digested with a proteolytic enzyme deposited with a dedicated device onto the tissue. After the proteolytic digestion, the selected MALDI matrix is sprayed onto the tissue and then the distribution of the Trastuzumab is determined based on its specific peptide mass fingerprint (PMF) by high resolution mass spectrometry imaging with a MALDI-FTICR instrument (SolariX FTICR 7.0T from Bruker). The images are obtained with Quantinetix software dedicated to the MSI data set.

Novel Aspect

We describe a process which combines MSI and bottom-up proteomics approach directly on tissue to follow the distribution of a therapeutic antibody without any labelling. Until now, mass spectrometry imaging was used to study some biomarkers' (proteins) distribution on tissue. This work describes a novel MSI application for analysis of a dosed therapeutic antibody. Data collection is ongoing.

Conclusions

We have adapted the use of Mass Spectrometry Imaging to follow the distribution of a non-labeled therapeutic antibody based on its peptide mass fingerprint in dosed tissues. This opens the use of this technology to other large molecules, such as therapeutic proteins or Antibody-Drug Conjugates (ADCs) in a variety of therapeutic fields (Oncology, Immunology, CNS, etc.)

In addition to the distribution of the target therapeutic antibody, Mass Spectrometry Imaging could also allow the detection of other important endogenous molecules in the same or adjacent tissue section, such as specific disease markers, thus allowing further understanding of PK/PD relationships.

Citation Format: David Bonnel, Chassidy Hall, Robert J. Mullin, Kathryn A. Simon, Jonathan Stauber. Mass Spectrometry Imaging of therapeutic antibodies: Distribution of unlabeled trastuzumab in CB.17 SCID mice implanted with the human breast BT474 xenograft. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2064. doi:10.1158/1538-7445.AM2014-2064