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.

 

 

  • doi:10.1681/ASN.2015050508
    “Increased Synthesis of Liver Erythropoietin with CKD”
    Author(s): Sophie de Seigneux, Anne-Kristine Meinild Lundby,Lena Berchtold, Anders H. Berg, Patrick Saudan and Carsten Lundby
  • doi:10.1002/dta.1916
    “Fc-fragment removal allows the EPO-Fc fusion protein to be detected in blood samples by IEF-PAGE”
    Author(s): P. Postnikov, G. Krotov, N. Mesonzhnik, Y. Efimova. G. Rodchenkov
    doi:10.1002/dta.1752
    “MAIIA EPO SeLect-a rapid screening kit for the detection of recombinant EPO analogues in doping control: inter-laboratory prevalidation and normative study of athlete urine and plasma samples”
    Author(s): Y. Dehnes L. Myrvold H. Ström M. Ericsson P. Hemmersbacha
  • doi:10.1371/journal.pone.0110903
    “Recombinant erythropoietin in humans has a prolonged effect on circulating erythropoietin isoform distribution”
    NJ. Aachmann-Andersen
    Author(s): S. Just Christensen K. Lisbjerg P. Oturai AK. Meinild-Lundby NH. Holstein-Rathlou C. Lundby N. Vidiendal Olsen
  • doi:10.1007/s00421-014-2844-7
    “Kidney-synthesized erythropoietin is the main source for the hypoxia-induced increase in plasma erythropoietin in adult humans”
    Author(s): AK. Lundby S. Keiser C. Siebenmann L. Schäffer C. Lundby
  • doi:10.1111/sms.12049
    “Detection of microdoses of rhEPO with the MAIIA test”
    Author(s): J. Mørkeberg K. Sharpe K. Karstoft M. J. Ashenden
  • doi:10.1039/C2AN15662H     Analyst. 137(10), 2445-2453 (2012).
    “A new analytical method based on anti-EPO monolith column and LC-FAIMS-MS/MS for the detection of rHuEPOs in horse plasma and urine samples”
    Author(s): Ludovic Bailly-Chouriberry Florence Cormant Patrice Garcia Maria Lönnberg Simon Szwandt Ulf Bondesson Marie-Agnès Popot Yves Bonnaire
  • doi:10.1007/s00216-012-5972-0 Anal. Bioanal. Chem. 403(6), 1619-1628 (2012).
    “Detection of recombinant human EPO administered to horses using MAIIA lateral flow isoform test”
    Author(s): Maria Lönnberg Ulf Bondesson Florence Cormant Patrice Garcia Yves Bonnaire Jan Carlsson Marie-Agnes Popot Niclas Rollborn Kristina Råsbo Ludovic Bailly-Chouriberry
  • doi:10.1111/j.1476-5381.2011.01822.x  Br. J. Pharmacol. 165(5), 1306-1315 (2012).
    “The evolving science of detection of ‘blood doping'”
    Author(s): Carsten Lundby Paul Robach Bengt Saltin
  • doi:10.1016/j.ab.2011.09.021  Anal. Biochem. 420(2), 101-114 (2012).
    “Rapid detection of erythropoiesis-stimulating agents in urine and serum”
    Author(s): Maria Lönnberg Maria Andrén Gunnar Birgegård Malin Drevin Mats Garle Jan Carlsson
  • doi:10.1007/s00216-011-5116-y Anal. Bioanal. Chem. 401(2), 463-481 (2011).
    “Recent developments in doping testing for erythropoietin”
    Author(s): Christian Reichel
  • doi:10.1016/j.jpba.2010.06.017 J. Pharm. Biomed. Anal. 53(4), 1028-1032 (2010).
    “Erythropoietin (EPO) immunoaffinity columns–a powerful tool for purifying EPO and its recombinant analogues”
    Author(s): Yvette Dehnes Severine Lamon Maria Lönnberg
  • doi:10.1016/j.chroma.2010.09.034 J. Chromatogr. A 1217, 7031-7037 (2010).
    “Rapid affinity purification of erythropoietin from biological samples using disposable monoliths”
    Author(s): Maria Lönnberg Yvette Dehnes Malin Drevin Mats Garle Severine Lamon Nicolas Leuenberger Trikien Quach Jan Carlsson
  • doi:10.1016/j.jim.2008.09.022 J. Immunol. Meth. 339, 236-244 (2008).
    “Ultra-sensitive immunochromatographic assay for quantitative determination of erythropoietin”
    Author(s): Maria Lönnberg Malin Drevin Jan Carlsson
  • doi:10.1016/j.chroma.2008.10.036 J. Chromatogr. A 1212, 82-88 (2008).
    “Lectin affinity chromatography as a tool to differentiate endogenous and recombinant erythropoietins”
    Author(s): Laura Franco Fraguas Jan Carlsson Maria Lönnberg
  • Bioforum Europe 11, 28-29 (2007).
    “Protein Isoform Determination”
    Author(s): Maria Lönnberg Torbjörn Karlsson Jan Carlsson
  • doi:10.1016/j.chroma.2006.06.016 J. Chrom. A 1127, 175-182 (2006).
    “Lab-on-a-chip technology for determination of protein isoform profiles”
    Author(s): Maria Lönnberg Jan Carlsson
  • Doctoral thesis, ISBN: 91-554-5250-7, Uppsala University, Sweden (2002).
    “Membrane-Assisted Isoform ImmunoAssay: Separation and determination of protein isoforms”
    Author(s): Maria Lönnberg
  • doi:10.1016/S0378-4347(01)00376-0 J. Chrom. B 763, 107-120 (2001).
    “Chromatographic performance of a thin microporous bed of nitrocellulose”
    Author(s): Maria Lönnberg Jan Carlsson
  • doi:10.1006/abio.2001.5130 Anal. Biochem. 293, 224-231 (2001).
    “Quantitative detection in the attomole range for immunochromatographic tests by means of a flatbed scanner”
    Author(s): Maria Lönnberg Jan Carlsson
  • doi:doi:10.1016/S0022-1759(00)00287-8 J. Immunol. Meth. 246, 25-36 (2000).
    “Membrane assisted isoform immunoassay: a rapid method for the separation and determination of protein isoforms in an integrated immunoassay”
    Author(s): Maria Lönnberg Jan Carlsson