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Resolution of Recombinant Human Interleukin 10 from Variants by Recycling Free Flow Focusing

Bondoc, Laureano L. ; Varnerin, Jeffrey P. ; Tang, John Chu-Tay

Analytical biochemistry, 1997-03, Vol.246 (2), p.234-238 [Periódico revisado por pares]

United States: Elsevier Inc

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  • Título:
    Resolution of Recombinant Human Interleukin 10 from Variants by Recycling Free Flow Focusing
  • Autor: Bondoc, Laureano L. ; Varnerin, Jeffrey P. ; Tang, John Chu-Tay
  • Assuntos: Humans ; Hydrogen-Ion Concentration ; Interleukin-10 - analysis ; Isoelectric Focusing - methods ; Recombinant Fusion Proteins - analysis
  • É parte de: Analytical biochemistry, 1997-03, Vol.246 (2), p.234-238
  • Notas: ObjectType-Article-1
    SourceType-Scholarly Journals-1
    ObjectType-Feature-2
    content type line 23
  • Descrição: Recombinant human interleukin 10 (rhIL-10) is a potential human therapeutic agent for treating inflammatory bowel diseases and rheumatoid arthritis. The rhIL-10 molecule derived fromEscherichia coliinclusion bodies consists of two identical subunits forming a noncovalent dimer. Since the ability to separate rhIL-10 from closely related impurities was highly desirable, recycling free flow focusing (RFFF) was utilized for the purification process development of rhIL-10. Under nondenaturing conditions, RFFF was able to separate rhIL-10 from fractions enriched in rhIL-10 variants. Three major monomeric variants (A, B, and C) can be identified and quantitated by reversed phase HPLC. The isoelectric point (pI) of rhIL-10 was empirically determined to be 8.2 while that for the three variant populations were in the range 7.3–7.5. Knowledge of these pI's would potentially facilitate the optimization process for ion-exchange chromatography. Furthermore, the technique provided a mild and fast preparation procedure for obtaining the recombinant protein and its variants for further characterization, as evidenced in the separation of rhIL-10 from variant C by successive RFFF treatments.
  • Editor: United States: Elsevier Inc
  • Idioma: Inglês
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