Metal chelate for protein adsorption studies pHEMA-GMA column filling materials including groups preparation

Year 2024, Volume: 13 Issue: 2, 522 - 527, 15.04.2024

Bahar Yılmaz Altınok , Mukaddes Keskinateş , Mevlüt Bayrakcı

https://doi.org/10.28948/ngumuh.1382364

Abstract

In this study, new poly(HEMA-co-GMA) or pHEMA-GMA support solid structures were prepared to help protein purification. The surface of the prepared pHEMA-GMA building materials was modified by binding different functional groups such as IDA and NTA. Nickel (Ni II) metal ion was passed through the modified cryogel structures. The monolithic cryogel structure chelated with Ni metal was characterized by SEM and FT-IR. GFP fluorescent protein binding studies of metal chelated cryogel structures were performed using UV-vis and fluorescence spectrophotometry. These cryogels were determined to bind 45% and 75% protein by UV-vis and fluorescence spectrophotometry. And these results were supported by visualization with the SDS PAGE method.

Keywords

p(HEMA-GMA), UV-vis and fluorescence spectrophotometer, SDS page, Nickel, IDA, NTA

Project Number

06-M-21

Protein adsorpsiyon çalışmaları için metal şelat grupları içeren pHEMA-GMA kolon dolgu malzemelerinin hazırlanması

Abstract

Bu çalışmada, protein saflaştırılmasına yardımcı olacak yeni poly(HEMA-co-GMA) veya pHEMA-GMA destek katı yapıları hazırlandı. Hazırlanan pHEMA-GMA yapı malzemelerin yüzeyi IDA ve NTA gibi farklı fonksiyonel grupların bağlanması ile modifiye edildi. Modifiye edilen kriyojel yapılarından nikel (Ni II) metal iyonu geçirildi. Ni metali ile şelatlanan monolitik kriyojel yapısı SEM ve FT-IR ile karakterize edildi. Metal şelatlı kriyojel yapıların GFP floresan protein bağlama çalışmaları UV-vis ve floresan spektrofotometresi ile yapıldı. Bu kriyojellerin UV-vis ve floresan spektrofotometresi ile %45 ve %75 oranında protein bağladıkları tespit edildi. Ve bu sonuçlar SDS PAGE yöntemi ile görselleştirilerek desteklendi.

Keywords

p(HEMA-GMA), UV-vis ve floresans spektrofotometresi, SDS page, Nikel, IDA, NTA

Ethical Statement

Yazarlar çıkar çatışması olmadığını beyan etmektedir.

Supporting Institution

Karamanoğlu Mehmetbey Üniversitesi, Bilimsel Araştırma Projeleri Komisyonu

Project Number

06-M-21

Thanks

06-M-21 numaralı projenin gerçekleştirilmesi için destek veren Karamanoğlu Mehmetbey Üniversitesi, Bilimsel Araştırma Projeleri Komisyonu’na teşekkür ederiz.

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