Naslov
Enzyme-linked immunosorbent assay for 4-hydroxynonenal-protein conjugates

Autori
Borović, Suzana ; Rabuzin, Filip ; Waeg, Georg ; Žarković, Neven

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
HNE-Club Meeting : Book of abstracts / - , 2006, 20-20

Skup
HNE-Club Meeting

Mjesto i datum
Genova, Italija, 16.06.2006. - 18.06.2006

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
4-hydroxynonenal; ELISA

Sažetak
Highly reactive aldehyde 4-hydroxynonenal (HNE) is the final product of lipid peroxidation, known as a second messenger of free radicals and a signaling molecule. It forms protein conjugates involved in pathology of various diseases. To determine cellular HNE– protein conjugates we developed indirect ELISA based on well-known, genuine monoclonal antibody against HNE– histidine (HNE– His) adducts. The method was calibrated using HNE– albumin conjugates as standards (R2 = 0.99) and validated on human osteosarcoma cell cultures (HOS). The ELISA showed good sensitivity (8.1 pmol HNE– His/mg of protein), precision (8% intra-assay and 12% inter-assay) and spiking recovery (9%). The assay revealed 60-fold increase of cellular HNE– His adducts upon copper-induced lipid peroxidation of HOS. The ELISA matched HNE-immunocytochemistry of HNE-treated HOS cells and quantified the increase of cellular HNE– His conjugates in parallel to the decrease of free HNE in culture medium. The ELISA was developed as ELISA Stress for severe lipid peroxidation and ELISA Fine for studies on HNE physiology.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

POVEZANOST RADA