engleski

IMMUNOAFFINITY HAPTOGLOBIN ISOLATION FROM HUMAN PLASMA USING MONOLITHIC SUPPORTS

Haptoglobin is positive acute phase protein, synthesized in the liver, with a reference range in adults 30 – 200 mg/dL. Its glycosylation, as one of the most significant post-translation modification, is altered in different types of disease. Many analytical techniques are described for glycosylation site and glycans analysis, but it is very important to isolate an optimal protein amount from plasma. The aim of this project was to obtain a protocol for haptoglobin isolation from human plasma for downstream haptoglobin glycosylation analysis. Protein isolation was based on principles of affinity chromatography. Different volumes of plasma samples and isolation conditions were tested to get more specific and effective isolation. Obtained protocol for haptoglobin isolation was applied on ten plasma samples from different male and female individuals. Glycans were analyzed by hydrophilic interaction ultra performance liquid chromatography (UPLC). Three haptoglobin phenotypes were present in samples and glycosylation varied between individuals.

haptoglobin, human plasma, affinity chromatography, monolithic supports

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