Jeroen Hans Nieuwenhuis

Rapid integrated biosensor for multiplexed immunoassays based on actuated magnetic nanoparticles

The realization of biomolecular detection assays for diagnostic purposes is technologically very challenging because such tests demand full integration for ease of use and need to deliver a high analytical performance with cost-effective use of materials. In this article an optomagnetic immunoassay technology is described based on nanoparticles that are magnetically actuated and optically detected in a stationary sample fluid. The dynamic control of nanoparticles by magnetic fields impacts the key immunoassay process steps, giving unprecedented speed, assay control and seamless integration of the total test. The optical detection yields sensitive and multiplexed assays in a low-cost disposable cartridge. We demonstrate that the optomagnetic technology enables high-sensitivity one-step assays in blood serum/plasma and whole saliva. Drugs of abuse are detected at sub-nanogram per millilitre levels in a total assay time of 1 min, and the cardiac marker troponin I is detected at sub-picomole per litre concentrations in a few minutes. The optomagnetic technology is fundamentally suited for high-performance integrated testing and is expected to open a new paradigm in biosensing.

Rapid, high sensitivity, point-of-care test for cardiac troponin based on optomagnetic biosensor

BACKGROUND We present a prototype handheld device based on a newly developed optomagnetic technology for the sensitive detection of cardiac troponin I (cTnI) in a finger-prick blood sample with a turnaround time of 5 min. METHODS The test was completed in a compact plastic disposable with on-board dry reagents and superparamagnetic nanoparticles. In our one-step assay, all reaction processes were precisely controlled using electromagnets positioned above and below the disposable. Nanoparticle labels (500 nm) bound to the sensor surface via a sandwich immunoassay were detected using the optical technique of frustrated total internal reflection. RESULTS A calibration function measured in plasma demonstrates a limit of detection (mean of blank plus 3-fold the standard deviation) of 0.03 ng/mL cTnI. A linear regression analysis of the region 0.03-6.5 ng/mL yields a slope of 37+/-4, and a linear correlation coefficient of R2=0.98. The measuring range could be extended substantially to 100 ng/mL by simultaneously imaging a second spot with a lower antibody concentration. CONCLUSIONS The combination of magnetic particles and their fine actuation with electromagnets permits the rapid and sensitive detection of cTnI. Because of the potential high analytical performance and ease-of-use of the test, it is well suited for demanding point-of-care diagnostic applications.

Confined Brownian motion of individual magnetic nanoparticles on a chip: Characterization of magnetic susceptibility.

An increasing number of biomedical applications requires detailed knowledge of the magnetic susceptibility of individual particles. With conventional techniques it is very difficult to analyze individual particles smaller than 1μm. The authors demonstrate how the susceptibility of individual nanoparticles can be determined in an efficient way by optically analyzing the confined Brownian motion of a nanoparticle trapped in a known magnetic potential well on a chip. A setup is introduced that has a controllable two-dimensional magnetic potential well, which is defined by an integrated microscopic current wire. Susceptibility measurements have been performed on 150–450nm superparamagnetic beads. They found differences in bead susceptibility of an order of magnitude and differences in volumetric susceptibility of more than a factor of 2.

Analysis of individual magnetic particle motion near a chip surface

We describe an analysis of the dynamics of individual superparamagnetic micro- and nanoparticles in order to quantify their magnetic properties and mobility near a chip surface. The particles are attracted to the chip surface by integrated microscopic current wires. We show that it is possible to accurately analyze particles with a diameter of about 1 μm by the magnetophoretic movement between current wires because of the very high field gradients. This reveals distinct differences in volume susceptibilities of particles with the same outer diameter. Smaller particles are characterized using the technique of confined Brownian motion analysis. By capturing 300 nm particles on a current wire with surface barriers or a focused shape, the magnetization of the particles can be measured with an accuracy better than 10%.

Analytical evaluation of a new point of care system for measuring cardiac Troponin I.

OBJECTIVES Point-of-care cardiac troponin testing with adequate analytical performances has the potential to improve chest pain patients flow in the emergency department. We present the analytical evaluation of the newly developed Philips Minicare cTnI point-of-care immunoassay. DESIGN & METHODS Li-heparin whole blood and plasma were used to perform analytical studies. The sample type comparison study was performed at 4 different hospitals. The 99th percentile upper reference limit (URL) study was performed using Li-heparin plasma, Li-heparin whole blood and capillary blood samples from 750 healthy adults, aging from 18 to 86years. RESULTS Limit of the blank, limit of detection and limit of quantitation at 20% coefficient of variation (CV) were determined to be 8.5ng/L, 18ng/L and 38ng/L respectively without significant differences between whole blood and plasma for LoQ. Cross-reactivity and interferences were minimal and no high-dose hook was observed. Total CV was found to be from 7.3% to 12% for cTnI concentrations between 109.6 and 6135.4ng/L. CV at the 99th percentile URL was 18.6%. The sample type comparison study between capillary blood, Li-heparin whole blood and Li-heparin plasma samples demonstrated correlation coefficients between 0.99 and 1.00 with slopes between 1.03 and 1.08. The method comparison between Minicare cTnI and Beckman Coulter Access, AccuTnI+3 demonstrated a correlation coefficient of 0.973 with a slope of 1.09. The 99th percentile URL of a healthy population was calculated to be 43ng/L with no significant difference between genders or sample types. CONCLUSIONS The Minicare cTnI assay is a sensitive and precise, clinical usable test for determination of cTnI concentration that can be used in a near-patient setting as an aid in the diagnosis of acute myocardial infarction.