MAIIA Technology and protein isoforms

MAIIA (Membrane Assisted Isoform ImmunoAssay) is a novel proprietary technology for rapid and sensitive measurement of protein isoforms in biological specimens. Posttranslational modifications of a protein, such as glycosylation, can significantly impact its biological functions. Despite numerous reports suggesting that protein isoform distributions produce clinical effects, there remains a lack of suitable methods for rapidly and reliably measuring protein isoforms, particularly ones that are present at low concentrations in blood and urine. MAIIA Technology has the ability to detect and resolve several types of post-translationally modified proteins at femto-molar concentrations.

MAIIA Technology is an affinity chromatography and an EPO immuno assay combined on a thin porous strip using three different tools for binding EPO: a PEG binding zone, an isoform separation zone based on lectin wheat germ agglutinin (WGA) and detection zone based on immobilized anti-EPO antibodies.  Capillary forces dictate the movement of the sample, from the sample application zone to subsequent separation, and capture zones. Pegylated substances such as CERA will be captured in the PEG binding zone, whereas all other EPO isoforms are first retained in the separation zone.  Bound EPO is then released by using a WGA competing sugar derivative N-acetylglucosamine (GlcNAc). The WGA ligand has different affinities with each isoform of EPO, resulting in different rates of migration for the various isoforms as they approach the detection zone. Urine and blood doped with recombinant EPOs, such as Aranesp, Neorecomon, CERA and EPO-FC, separate differently on the MAIIA-strips and can, therefore, be used for screening in anti-doping applications.

EPO that is bound to the detection zone is visualized with Anti-EPO Carbon Black Nano-Strings (Anti-EPO CBNS) in a sandwich configuration yielding a grey to black signal. The black intensity is proportional to the amount of bound EPO and is converted into an empirical value via a flatbed scanner and image processing software, developed by MAIIA Diagnostics. The detection limit can reach as low as 1.2 femtomolar or 0.035 ng EPO/L.



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