ISSN: 2155-9872

Zeitschrift für analytische und bioanalytische Techniken

Offener Zugang

Unsere Gruppe organisiert über 3000 globale Konferenzreihen Jährliche Veranstaltungen in den USA, Europa und anderen Ländern. Asien mit Unterstützung von 1000 weiteren wissenschaftlichen Gesellschaften und veröffentlicht über 700 Open Access Zeitschriften, die über 50.000 bedeutende Persönlichkeiten und renommierte Wissenschaftler als Redaktionsmitglieder enthalten.

Open-Access-Zeitschriften gewinnen mehr Leser und Zitierungen
700 Zeitschriften und 15.000.000 Leser Jede Zeitschrift erhält mehr als 25.000 Leser

Indiziert in
  • CAS-Quellenindex (CASSI)
  • Index Copernicus
  • Google Scholar
  • Sherpa Romeo
  • Datenbank für wissenschaftliche Zeitschriften
  • Öffnen Sie das J-Tor
  • Genamics JournalSeek
  • JournalTOCs
  • Forschungsbibel
  • Nationale Wissensinfrastruktur Chinas (CNKI)
  • Ulrichs Zeitschriftenverzeichnis
  • Elektronische Zeitschriftenbibliothek
  • RefSeek
  • Verzeichnis der Indexierung von Forschungszeitschriften (DRJI)
  • Hamdard-Universität
  • EBSCO AZ
  • OCLC – WorldCat
  • Gelehrter
  • SWB Online-Katalog
  • Virtuelle Bibliothek für Biologie (vifabio)
  • Publons
  • Euro-Pub
  • ICMJE
Teile diese Seite

Abstrakt

Optimal Concentration of 2,2,2-Trichloroacetic Acid for Protein Precipitation Based on Response Surface Methodology

Albert N Ngo, Miezan JM Ezoulin, Ibrahima Youm and Bi-Botti C Youan

For low protein concentrations containing biological samples (in proteomics) and for non proteinaceous compound assays (in bioanalysis), there is a critical need for a simple, fast, and cost-effective protein enrichment or precipitation method. However, 2,2,2-trichloroacetic acid (TCA) is traditionally used for protein precipitation at ineffective concentrations for very low protein containing samples. It is hypothesized that response surface methodology, can be used to systematically identify the optimal TCA concentration for protein precipitation in a wider concentration range. To test this hypothesis, a central composite design is used to assess the effects of two factors (X1 = volume of aqueous solution of protein, and X2 = volume of TCA solution 6.1N) on the optical absorbance of the supernatant (Y1), and the percentage of protein precipitated (Y2). Using either bovine serum albumin (BSA) as a model protein or human urine (with 20 ppm protein content), 4% w/v (a saddle point) is the optimal concentration of the TCA solution for protein precipitation that is visualized by SDS-PAGE analysis. At this optimal concentration, the Y2-values range from 76.26 to 92.67% w/w for 0.016 to 2 mg/mL of BSA solution. It is also useful for protein enrichment and xenobiotic analysis in protein-free supernatant as applied to tenofovir (a model HIV microbicide). In these conditions, the limit of detection and limit of quantitation of tenofovir are respectively 0.0014 mg/mL and 0.0042 mg/mL. This optimal concentration of TCA provides optimal condition for protein purification and analysis of any xenobiotic compound like tenofovir.