Naslov (eng)

Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum-A New Biotech Solution

Autor

Četić, Danilo
Miljuš, Goran
Dobrijević, Zorana
Gligorijević, Nikola
Vilotić, Aleksandra
Nedić, Olgica
Penezić, Ana

Publisher

Opis (eng)

ABSTRACT A fast and simple biotech method is presented for the simultaneous isolation and purification of transferrin (Tf) and immunoglobulin G (IgG) from the same pool-sample of human serum, yielding >98% pure proteins. Serum sample preparation was achieved by precipitation with ethacridine lactate (rivanol). Protein purification was performed with AKTA Avant 150 FPLC, using a Resource Q column. Three different buffers at pH 6.2 (MES, phosphate, and Bis-Tris) were tested. Isolated and purified proteins retained their native 3D structure, as shown by spectrofluorimetric measurements. Tf functionality was preserved, as confirmed by the retention of both the iron binding capacity and its ability to interact with the transferrin receptor (immunofluorescent staining), as well as the immunogenicity of IgG, as shown by Western blot analysis with immunodetection. The formation of IgG aggregates was avoided. This biotech method is a rapid, simple, and time-saving alternative to other methods for the isolation of extremely pure IgG and Tf, while it is also the only method so far described for their simultaneous isolation.

Jezik

engleski

Datum

2025

Licenca

Ovo delo je licencirano pod uslovima licence
Creative Commons CC BY 4.0 - Creative Commons Autorstvo 4.0 International License.

http://creativecommons.org/licenses/by/4.0/legalcode

Predmet

biotherapeutics; isolation and purification; ion-exchange; IgG; transferrin

Deo kolekcije (1)

o:25140

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