Development of Analytical Techniques for Biological Marker Analysis

ISBN-10
1109960786
ISBN-13
9781109960785
Category
Biochemical markers
Pages
164
Language
English
Published
2007
Author
Matthew Michael Petkus

Description

Rising interest in how diseases evolve through the monitoring of biological markers has created the need for enhanced sensing techniques. Immunoassay and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) techniques have been altered for the improved detection and quantification of markers in micro liter volume samples. Fluoroimmunoassays have been developed using paramagnetic particles as solid phases. Through binding antibodies to micron diameter iron oxide particles, antigen is rapidly captured through convective mixing. Introduction of a magnetic field creates dipole moments on the particles, attracting them to form rod-like supraparticle structures. These structures can rotate within an alternating magnetic field generating a periodic signal that can be analyzed with lock-in amplification allowing more sensitive detection. Using this modulated immunoassay fluorescently labeled cortisol, a hormone linked to stress, has been quantified from 300pM--30nM, a range useful for saliva analysis. A modulated sandwich assay has also quantified myoglobin, a marker for acute myocardial infarction, in serum samples from 50pM--1nM. A modulated competitive assay demonstrated the capability of rapidly detecting myoglobin concentrations greater than or equal to 5 nM. Detection of myoglobin using MALDI-TOF MS and a grapho epitaxy approach has also been investigated. The grapho epitaxy approach uses lithographically patterned silica in an attempt to control nucleation and crystallization of mixtures of sinipic acid and myoglobin. The effect of crystal formation was investigated using different temperature, matrix concentrations, and evaporative rates.

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