Bruker Corporation and SISCAPA Assay Technologies, Inc. have announced the publication of a study in the Journal of Proteome Research demonstrating that SISCAPA MALDI-TOF mass spectrometry provides excellent precision for quantitation of proteins of interest, such as high value protein biomarkers.
The publication results from an ongoing collaboration between Bruker and SAT to develop MALDI-TOF as a high precision and high throughput mass spectrometric detection method for SISCAPA (Stable Isotope Standards and Capture by Anti-Peptide Antibodies) assays.
The collaboration aims to exploit the high throughput, robustness and ease of use of MALDI-TOF instruments as an alternative to nano-LC-MS technology currently used in many SISCAPA assays.
While the ease of use and robustness of MALDI-TOF make it attractive for high throughput biomarker validation and pre-clinical studies, MALDI-TOF has generally been considered to have insufficient quantitative precision to be used in most clinical biomarker assays.
In recent years, however, MALDI-TOF technology has advanced dramatically with high repetition rate lasers (up to 1 kHz) and automated self cleaning ion sources, resulting in faster, 24/7 operation with consistent results.
As shown in the paper published in the Journal of Proteome Research, the use of internal, isotopically labeled peptide standards in the SISCAPA assay yields very precise relative quantitation of peptides (CVs of 1-2%), comparing favorably with results obtained by the best conventional nano-LC-MS workflows.
The elimination of complex and time-consuming nano-flow LC-MS separation greatly decreases the complexity of sample handling and sample introduction, which, together with advances in automated preparation of MALDI target plates, enables large scale biomarker verification studies currently considered impractical.
The published results were obtained using a high- performance Bruker autoflex speed MALDI-TOF mass spectrometer, and unpublished results further indicate that assays for medium and high abundance proteins also are possible on the benchtop microflex MALDI-TOF used in Bruker’s MALDI Biotyper solution in clinical microbiology, and aimed at clinical and GMP environments.
Leigh Anderson, Ph.D., the CEO and founder of SISCAPA Assay Technologies, and inventor of SISCAPA, commented: “We are delighted to be working with Bruker, the leader in MALDI-TOF technology, to advance the precision and throughput of SISCAPA biomarker assays. Our joint paper demonstrates the very high precision of MALDI-TOF for peptide quantitation using internal standards, and establishes a solid basis for further exploring the utility of SISCAPA MALDI-TOF in biomarker verification and preclinical research. As a company devoted to the development and application of SISCAPA assays, we are excited that the addition of this robust and easy MALDI-TOF workflow to SISCAPA will not only increase the productivity of labs using SISCAPA, but also increase the adoption of SISCAPA assays by the biomarker research community.”
Dr. Detlev Suckau, Director of Proteomics at Bruker Daltonics, added: “The results of the joint work prove that MALDI-TOF is very suitable for peptide and protein quantification in the 1-5 % CV range, if done properly. This is a very positive development, as previously many proteomics researchers may have considered MALDI-TOF as primarily a qualitative or semi-quantitative platform. We are very pleased to continue to work with Leigh Anderson and SISCAPA Assay Technologies on these ground-breaking biomarker quantification and validation developments. The results in this publication suggest that the use of the ‘LC-free’ SISCAPA-MALDI-TOF workflow in pre-clinical research is quite attractive. This is an emerging market for Bruker that will further increase the visibility of our very successful MALDI-TOF product line in biomedical research.”