Prostate-specific antigen (PSA) glycan-binding profile analysis based on enzyme-linked lectin assay (ELLA) and storage effect of assay components

Nur Hanina Izzati Khairol Mokhtar; Goh Dirong; Muhammad Ashraf Shahidan

Authors

Nur Hanina Izzati Khairol Mokhtar Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, The National University of Malaysia, 43600 UKM Bangi, Selangor D.E., Malaysia
Goh Dirong Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, The National University of Malaysia, 43600 UKM Bangi, Selangor D.E., Malaysia
Muhammad Ashraf Shahidan Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, The National University of Malaysia, 43600 UKM Bangi, Selangor D.E., Malaysia

Keywords:

glycoprofiling, lectin assay, MAA, PSA, sialic acid, SNA, storage

Abstract

Changes in the prostate-specific antigen (PSA) glycosylation profile may be used to distinguish indolent from aggressive prostate cancer (PCa). This study aimed to obtain the glycan profiles of PSA isolated from a normal individual using the enzyme-linked lectin assay (ELLA) method. We used sialic acid-specific lectins (MAA I, MAA II and SNA) for PSA glycoprofiling and generated the binding curve of PSA-lectin interactions. Compared to MAA I and MAA II, Sambucus nigra agglutinin (SNA) had the highest binding (A₄₅₀ = 3.59) with the normal individual PSA sample harbouring α2,6-sialic acid glycan used in this study. The binding signals for Maackia amurensis agglutinin (MAA) I were significant starting at 700 ng/mL PSA, implying that a small amount of α2,3-sialic acid glycan presented in the normal PSA. Binding saturation occurred at A₄₅₀ = 3.60 and at a PSA concentration of 5 × 10³ ng/mL (176 nM). In addition, the stability of assay components stored at room temperature (25 °C) for two months was assessed, and the results showed that the binding signals for PSA detection by SNA were still remarkably high (A₄₅₀ = 2.37) at a PSA concentration of 1 µg/mL. Here, we established a simple and sensitive lectin-based assay for PSA glycoprofiling. The stability of the assay components during the storage test also revealed their potential to be utilised and stored for longer periods at room temperature.

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