Development of Nanoparticle-Based Mass Spectrometric Immunoassay for Targeted Proteomics
Date Issued
2005
Date
2005
Author(s)
Chou, Po-Hung
DOI
en-US
Abstract
As one of the most complex proteomes, human serum provides rich informative molecules for diagnostic analyses because the regulation and expression of proteins (putative biomarkers) into the circulating bloodstream are correlated to specific physiological states. Serum contains thousands of proteins, but most abundant twenty-two proteins constitute ~99 % of the protein content. The depletion of high-abundance proteins such as albumin or immunoglobulin is usually an essential yet time-consuming step for low-abundance protein profiling.
Alternatively, the combination of immunoassay with mass spectrometry has the potential to detect, isolate, and identify a vast range of target analytes in complex mixtures. Magnetic nanoparticles (mNP) immobilized with antibodies on their surfaces have a promising potential in isolation and preconcentration of targeted proteins. In addition to their comparable dimensions to the protein, the intrinsic magnetic property makes them easy to be manipulated by an external magnetic field for protein purifications.
In this thesis, the surface of mNP was covalently encapsulated with specific antibodies which served as “bait molecules” to recognize and isolate disease-related antigens, such as CRP (C-reactive protein) and SAP (Serum amyloid P component) from normal and diseased human sera. The immunoaffinity-captured processes would combine with the power of unambiguous, rapid, sensitive, and “direct” protein identification of MALDI MS. Low abundance proteins in subnanomolar (10-9 ~10-10 M) was successfully detected. When it was applied to screen the CRP and SAP expression profiles of healthy and patient serum suffered from gastric cancer, there was a good consistency between this approach and conventional ELISA readouts.
Subjects
急性期蛋白
基質輔助雷射脫附游離質譜法
磁性奈米粒子
親和質譜術
MALDI MS
magnetic nanoparicles
affinity mass spectrometry
acute-phase proteins
SDGs
Type
thesis
File(s)![Thumbnail Image]()
Loading...
Name
ntu-94-R91223052-1.pdf
Size
23.31 KB
Format
Adobe PDF
Checksum
(MD5):7aed349a2faf0d62d69675110beefa11