Nedelkov D.,Intrinsic Bioprobes Inc. |
Nedelkov D.,Institute for Population Proteomics
Expert Review of Molecular Diagnostics | Year: 2012
Mass spectrometry-based protein assays hold great promise for in vitro diagnostic testing. Technological advances in mass spectrometry have given rise to instruments and methods that are fully capable of automated and high-throughput protein assaying. Yet, the numerous steps involved in such assays can lead to difficulties in assay characterization and validation, and can also make them unnecessarily complex and prohibitively expensive for everyday use. Simplification of both approaches and instrumentation seems to be the solution to the fast introduction of the mass spectrometry-based assays into the clinical laboratories. One such simplified approach is the mass spectrometric immunoassay, which couples targeted immunoaffinity protein separation with the power of mass spectrometry detection. Several mass spectrometric immunoassays have been extensively characterized and have found their way into clinical laboratory improvement amendments-certified laboratories in the form of laboratory developed tests. Reviewed in this special report is the development and validation of one of those assays - a Cystatin mass spectrometric immunoassay. With the added advantage of protein variant detection and quantification, these assays can redefine our view of protein diversity, with clear implications in biomarker discovery, validation, and ultimately, in vitro diagnostic testing. © 2012 Expert Reviews Ltd. Source
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 373.30K | Year: 2011
This proposal sets forth a precise and rapid methodology for identifying the interaction sites between monoclonal antibodies and proteins. The basis of the system is a high-throughput mass spectrometry-based analysis of epitope determinants to identify theinteracting protein sequences using immunoaffinity capture combined with enzymatic digestion of the target protein; either pre- or post-capture. This combination will be used to rapidly determine the approximate location of each binding epitope. The precise location of the binding epitope will then be determined by immunoaffinity capture and mass spectrometric readout of sets of synthetic ladder peptides that span the approximate epitope established from the initial analysis. The project will develop reproducible methods to demonstrate epitope mapping for monoclonal antibodies of interest to NCI tested by multiple sampling and quantifiable results. Mapped epitopes will be functionally characterized by amino acid substitutions in the epitope sequence using high-throughput spotted array surface plasmon resonance imaging and SPR-Biomolecular Interaction Analysis to identify the key amino acids that are critically involved in binding the antibody to the antigenic epitope. Thus, this approach will provide an efficient and cost effective platform from which to accomplish high-throughput epitope mapping of monoclonal antibodies for use in diagnosis of cancer.
Nedelkov D.,Intrinsic Bioprobes Inc.
Methods in molecular biology (Clifton, N.J.) | Year: 2010
The combination of surface plasmon resonance (SPR) and mass spectrometry (MS) creates a comprehensive protein investigation approach wherein SPR is employed for protein quantification and MS is utilized to structurally characterize the proteins. In such, MS utterly complements the SPR detection and reveals intrinsic protein structural modifications that go unregistered via the SPR detection. Protein complexes and non-specific binding can also be delineated via the SPR-MS approach. Described here are the protocols and know-how for successful and reproducible integration of SPR and MS. The individual steps of the entire SPR-MS process are illustrated via an example showing analysis of myoglobin from human plasma. Source
Intrinsic Bioprobes Inc. | Date: 2013-02-11
Described is an affinity microcolumn comprising a high surface area material, which has high flow properties and a low dead volume, contained within a housing and having affinity reagents bound to the surface of the high surface area material that are either activated or activatable. The affinity reagents bound to the surface of the affinity microcolumn further comprise affinity receptors for the integration into high throughput analysis of biomolecules.
Intrinsic Bioprobes Inc. | Date: 2015-01-30
The present invention is directed to diagnosing, determining, and/or monitoring type 2 diabetes, pre-diabetes, insulin resistance, and their misted conditions by detecting levels and modulations of ApoCIII and its variants. The present invention is also directed to methods for identifying and evaluating therapeutic treatments for type 2 diabetes, pre-diabetes, insulin resistance, and their related conditions by monitoring ApoCIII and its variants.