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SRM/MRM assay for the mesothelin (MSLN) protein

一种蛋白质、皮素的技术，应用在测量装置、生物测试、化学分析中使用的标记等方向

Active Publication Date: 2018-03-27
EXPRESSION PATHOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these agents also kill growing normal cells, so these agents are not considered a "targeted" method of killing cancer cells

Method used

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  • SRM/MRM assay for the mesothelin (MSLN) protein

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Embodiment Construction

[0014] In principle, any predicted peptide from an MSLN protein, e.g. prepared by digestion with a known specific protease (e.g., trypsin), can be used as a surrogate reporter for use with mass spectrometry-based SRM / MRM assay to determine the abundance of MSLN protein in the sample. Similarly, it is also possible to use any predicted peptide sequence that contains an amino acid residue at a site known to be likely to be modified in the MSLN protein to determine the degree of modification of the MSLN protein in a sample.

[0015]MSLN fragment peptides can be produced by a variety of methods, including using the Liquid Tissue protocol provided in US Patent 7,473,532. Liquid Tissue protocols and reagents enable the generation of mass spectrometry-ready peptide samples from formalin-fixed, paraffin-embedded tissues by proteolytic digestion of proteins in tissue / biological samples. In the Liquid Tissue protocol, the tissue / biological preparation is heated in buffer for an extende...

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Abstract

The current disclosure provides methods for quantifying the MSLN protein directly in biological samples, including samples that have been fixed in formalin by Selected Reaction Monitoring / Multiple Reaction Monitoring (SRM / MRM) mass spectrometry. Samples that can be assayed include tissues and cells treated with formaldehyde containing agents / fixatives including formalin-fixed tissue / cells, formalin-fixed / paraffin embedded (FFPE) tissue / cells, FFPE tissue blocks and cells from those blocks, and tissue culture cells that have been formalin fixed and or paraffin embedded. Methods to prepare a protein sample from such a biological sample are provided and MSLN protein is quantitated by SRM / MRM mass spectrometry by quantitating in the protein sample at least one or more of the peptides described.

Description

[0001] This application claims priority to Provisional Application No. 62 / 161,506, filed May 14, 2015, the entire contents of which are incorporated herein by reference. Background technique [0002] Cancer is treated with a panel of therapeutic agents that kill growing and dividing cells and function in a variety of ways. A common set of chemotherapeutic agents has been used alone or in combination for decades, and this common set of agents has become a traditional and routine cancer treatment in clinical oncology practice. These traditional chemotherapy agents work by killing all rapidly dividing cells, one of the main properties of most cancer cells. However, these agents also kill growing normal cells, so these agents are not considered a "targeted" method of killing cancer cells. In recent years, a large number of cancer therapeutic agents that selectively target only cancer cells have been developed, wherein these therapeutic agents specifically attack proteins expresse...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/574G01N33/6848G01N2333/47G01N2333/976G01N2800/56G01N2800/7028G01N33/6851G01N2458/15
Inventor 大卫·B·克里茨曼托德·哈姆布拉夫阿黛尔·布兰克勒史诺·塞帕洛姆比尔安恩京
Owner EXPRESSION PATHOLOGY
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