Capillary electrophoresis for simultaneous quantification of human proinsulin, insulin and intermediate forms
Articolo
Data di Pubblicazione:
1998
Citazione:
Capillary electrophoresis for simultaneous quantification
of human proinsulin, insulin and intermediate forms / C.Arcelloni, L. Falqui, S. Martinenghi, A.E. Pontiroli, R. Paroni. - In: ELECTROPHORESIS. - ISSN 0173-0835. - 19:8-9(1998 Jun), pp. 1475-1477.
Abstract:
Capillary electrophoresis (CE) for the simultaneous and precise quantification
of human insulin (hI), proinsulin (hPI) and intermediate forms (des 31, 32;
split 65-66 and des 64, 65), released in culture media by engineered cells, is
described. Analytical conditions for standard proteins were optimized using a
bare silica capillary (20 cm X 50 pm internal diameter). Proteins were monitored
at 200 nm and separated at constant voitage. Culture supernatants
(12-24 mL) were purified on Sep-Pak Vac C,* cartridges, recovered in 1 mL of
acetonitrile:trifluoracetic acid mixture (60:40, v:v), concentrated, ultrafiltered and injected into CE. Protein recovery was 85 + 140/o (n:5, mean -l- standard deviation) with a sensitivity limit of 0.5 nmol/L in the culture media, corresponding
to 2 fmol injected in22 nL. Using the CE method, it was possible to
detect and quantify, with precision and accuracy, the release of hPI, hI and
intermediate forms directly in the cell culture media, and to compare the proteic pattern released from engineered cells transduced with different hPI gene
constructs.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
Capillary electrophoresis; Proinsulin conversion; Intermediate
forms; Culture media purification
Elenco autori:
C.Arcelloni, L. Falqui, S. Martinenghi, A.E. Pontiroli, R. Paroni
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