Magnetic immunoassay
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Magnetic immunoassay (MIA) is a type of diagnostic immunoassay using magnetic beads as labels in lieu of conventional enzymes (ELISA), radioisotopes (RIA) or fluorescent moieties (fluorescent immunoassays)[1] to detect a specified analyte. MIA involves the specific binding of an antibody to its antigen, where a magnetic label is conjugated to one element of the pair. The presence of magnetic beads is then detected by a magnetic reader (magnetometer) which measures the magnetic field change induced by the beads. The signal measured by the magnetometer is proportional to the analyte (virus, toxin, bacteria, cardiac marker, etc.) concentration in the initial sample.
Magnetic beads are made of nanometric-sized iron oxide particles encapsulated or glued together with polymers. These magnetic beads range from 35 nm up to 4.5 μm. The component magnetic nanoparticles range from 5 to 50 nm and exhibit a unique quality referred to as superparamagnetism in the presence of an externally applied magnetic field.[2] First discovered by Frenchman Louis Néel, Nobel Physics Prize winner in 1970, this superparamagnetic quality has already been used for medical application in Magnetic Resonance Imaging (MRI) and in biological separations, but not yet for labeling in commercial diagnostic applications. Magnetic labels exhibit several features very well adapted for such applications:[citation needed]
- they are not affected by reagent chemistry or photo-bleaching and are therefore stable over time,
- the magnetic background in a biomolecular sample is usually insignificant,
- sample turbidity or staining have no impact on magnetic properties,
- magnetic beads can be manipulated remotely by magnetism.
Detection
Magnetic Immunoassay (MIA) is able to detect select molecules or pathogens through the use of a magnetically tagged antibody. Functioning in a way similar to that of an ELISA or Western Blot, a two-antibody binding process is used to determine concentrations of analytes. MIA uses antibodies that are coating a magnetic bead. These anti-bodies directly bind to the desired pathogen or molecule and the magnetic signal given off the bound beads is read using a magnetometer. The largest benefit this technology provides for immunostaining is that it can be conducted in a liquid medium, where methods such as ELISA or Western Blotting require a stationary medium for the desired target to bind to before the secondary antibody (such as HRP [Horse Radish Peroxidase]) is able to be applied. Since MIA can be conducted in a liquid medium a more accurate measurement of desired molecules can be performed in the model system. Since no isolation must occur to achieve quantifiable results users can monitor activity within a system. Getting a better idea of the behavior of their target. [citation needed]
The manners in which this detection can occur are very numerous. The most basic form of detection is to run a sample through a gravity column that contains a polyethylene matrix with the secondary anti-body. The target compound binds to the antibody contained in the matrix, and any residual substances are washed out using a chosen buffer. The magnetic antibodies are then passed through the same column and after an incubation period, any unbound antibodies are washed out using the same method as before. The reading obtained from the magnetic beads bound to the target which is captured by the antibodies on the membrane is used to quantify the target compound in solution.[citation needed]
Also, because it is so similar in methodology to ELISA or Western Blot the experiments for MIA can be adapted to use the same detection if the researcher wants to quantify their data in a similar manner.