Mass spectrometric immunoassay analysis of specific proteins and variants present in various biological fluids
US7396687B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Oct 4, 2004 |
| Grant date | Jul 8, 2008 |
| Priority date | — |
| Expiry date | Oct 4, 2024 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10T436/24
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Presented herein is the construction of pipettor tips (termed MSIA-Tips) containing porous solid supports that are constructed, covalently derivatized with affinity ligand, and used to extract specific proteins and their variants from various biological fluids by repeatedly flowing the fluids through the MSIA-Tips. A second protein species (a mass-shifted variant of the targeted protein doped into the samples at a constant concentration) is co-extracted with the endogenous protein and variants and is used as a quantitative internal reference standard (IRS). Nonspecifcally bound compounds are rinsed from the MSIA-Tip using a series of buffer and water rinses, after which the wild type protein, protein variants and the IRS are eluted from the MSIA-Tips directly onto a target in preparation for analysis such as MALDI-TOF. Mass spectrometry of the eluted sample then follows with the retained proteins identified via accurate molecular mass determination. Protein and variant levels are determined via a quantitative method in which the protein/variant signals are normalized to the signal of the IRS and the values compared to a working curve constructed from samples containing known concen…
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