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J Immunoassay Immunochem ; 34(2): 180-96, 2013.
Article in English | MEDLINE | ID: mdl-23537302

ABSTRACT

Adiponectin (Adn) is a protein that circulates in the blood in several oligomeric forms, namely low-, medium-, and high-molecular-weight forms. Adn may serve as a risk factor for type 2 diabetes mellitus (T2DM). The aims of this work were (1) to produce monoclonal antibodies (MAbs) specific to different Adn oligomeric forms, (2) to design immunoassays suitable for measuring the Adn forms present in human blood, and (3) to investigate the changes in Adn forms that occur in patients with T2DM. Gel filtration, fluoroimmunoassays, and Western blotting were utilized as major techniques in this study. MAbs recognizing various oligomeric forms of Adn were obtained. Complexes between Adn and complement component C1q and between the low molecular weight form of Adn and albumin were described in human blood. A decrease in the total Adn and Adn-albumin complex levels in the blood of patients with T2DM and no difference in the levels of the Adn-C1q complex in comparison with healthy volunteers were demonstrated. An Adn94-Adn63 fluoroimmunoassay was selected as the technique that most accurately measured the mass ratio of Adn oligomers in blood samples, and an Adn214-Adn27 assay that measured the low-molecular-weight form of Adn only.


Subject(s)
Adiponectin/blood , Diabetes Mellitus, Type 2/blood , Immunoconjugates/blood , Adiponectin/chemistry , Adult , Aged , Animals , Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/chemistry , Case-Control Studies , Complement C1q/chemistry , Female , Humans , Immunoconjugates/chemistry , Male , Mice , Middle Aged , Molecular Weight , Protein Binding , Protein Isoforms/blood , Protein Isoforms/chemistry , Serum Albumin/chemistry
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