SUMMARYIn this paper, we propose improved methods of liquidphase detection of biological targets utilizing magnetic markers and a high-critical-temperature superconducting quantum interference device (SQUID). For liquid-phase detection, the bound and unbound (free) markers are magnetically distinguished by using Brownian relaxation of free markers. Although a signal from the free markers is zero in an ideal case, it exists in a real sample on account of the aggregation and precipitation of free markers. This signal is called a blank signal, and it degrades the sensitivity of target detection. To solve this problem, we propose improved detection methods. First, we introduce a reaction field, B re , during the binding reaction between the markers and targets. We additionally introduce a dispersion process after magnetization of the bound markers. Using these methods, we can obtain a strong signal from the bound markers without increasing the aggregation of the free markers. Next, we introduce a field-reversal method in the measurement procedure to differentiate the signal from the markers in suspension from that of the precipitated markers. Using this procedure, we can eliminate the signal from the precipitated markers. Then, we detect biotin molecules by using these methods. In an experiment, the biotins were immobilized on the surfaces of large polymer beads with diameters of 3.3 μm. They were detected with streptavidinconjugated magnetic markers. The minimum detectable molecular number concentration was 1.8 × 10 −19 mol/ml, which indicates the high sensitivity of the proposed method. key words: high-critical-temperature superconducting quantum interference device (SQUID), magnetic marker, immunoassays, liquid-phase detection

We developed a new and improved method for the liquid-phase detection of biological targets using magnetic markers. Unlike conventional studies, we performed a binding reaction between markers and targets in the presence of a magnetic field. This field acts to prevent the Brownian rotation of markers during the reaction. In this case, markers are bound to the targets with their magnetic moments (m’s) aligned, which is in contrast to the conventional case where m’s are randomly oriented after the reaction. As a result, we could obtain much larger signals from the bound markers without increasing the blank signal from the free markers.

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