Silica microspheres functionalized with the iminodiacetic acid/copper(II) complex as a peroxidase mimic for use in metal affinity chromatography-based colorimetric determination of histidine-tagged proteins

Gokcal B., Kip C., Sahinbas D., Celik E., Tuncel A.

MICROCHIMICA ACTA, cilt.187, sa.2, 2020 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 187 Sayı: 2
  • Basım Tarihi: 2020
  • Doi Numarası: 10.1007/s00604-019-4087-0
  • Dergi Adı: MICROCHIMICA ACTA
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Aerospace Database, Analytical Abstracts, Aqualine, Aquatic Science & Fisheries Abstracts (ASFA), Biotechnology Research Abstracts, Chemical Abstracts Core, Chimica, Communication Abstracts, EMBASE, Food Science & Technology Abstracts, MEDLINE, Metadex, Pollution Abstracts, Civil Engineering Abstracts
  • Anahtar Kelimeler: Protein assay, Nanozyme, Peroxidase-like activity, Hydrogen peroxide, Green fluorescent protein, SiO2, Adsorption, Fluoresence, Iminodiacetic acid, Metal chelate complex, CATALYTIC-ACTIVITY, SENSITIVE DETECTION, PURIFICATION, FRAMEWORK, IMMOBILIZATION, NANOCOMPOSITE, NANOPARTICLES, FLUORESCENCE, INHIBITION, COPPER(II)
  • Hacettepe Üniversitesi Adresli: Evet

Özet

Monodisperse porous silica microspheres were functionalized with the iminodiacetic acid/copper(II) complex and then evaluated as a group-specific peroxidase-mimicking nanozyme for colorimetric determination of histidine-tagged (His-tagged) proteins. The green fluorescent protein (GFP) was selected as a typical His-tagged protein. The specificity for GFP and the peroxidase-like activity for the selected substrate were obtained by immobilizing the complex on the porous microspheres. The modified microspheres were also evaluated as a group specific immobilized metal affinity chromatography (IMAC) sorbent for the purification of GFP from Escherichia coli extract. The peroxidase-like activity of the microspheres was inhibited by the GFP adsorbed onto the microspheres due to the interaction of His-tagged protein with the immobilized Cu(II) complex. Ortho-phenylenediamine is used as a substrate for the enzyme mimic. The photometric response (measured at 416 nm) is linear in the 9.0-92 mu g center dot mL(-1) GFP concentration range in E. coli lysate. The limit of detection is 6.9 mu g center dot mL(-1).