Naslov
Use of short monolithic columns for isolation of low abundance membrane proteins

Autori
Ručević, Marijana ; Clifton, James G. ; Huangb, Feilei ; Li, Xuesong ; Callanan, Helen ; Hixson, Douglas C. ; Josić, Đuro

Izvornik
Journal of Chromatography A (0021-9673) 1123 (2006), 2; 199-204

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
plasma membrane proteins; immunoaffinity chromatography; monoliths

Sažetak
Convective interaction media (CIM) monoliths provide a stationary phase with a high binding capacity for large molecules and are capable of high flow rates at a very low pressure drop. Used as anion- and cation-exchangers or with affinity ligands such as antibodies, these columns have the potential for processing large volumes of complex biological mixtures within a short time. In the present report, monoclonal antibodies against several rat liver plasma membrane proteins were bound and cross-linked to protein A or protein G CIM affinity columns with a bed volume of only 60
L. Antigens recognized by bound antibodies and co-eluting (interacting) proteins were rapidly isolated in a single step from either total plasma membrane extracts or subfractions isolated using anion-exchange CIM disk-shaped columns. The isolated antigens and co-eluting proteins were subsequently identified by immunoblot or by LC– MS/MS.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija

Napomena
Rad ostvaren u suradnji s: Division of Medical Oncology, Department of Medicine, Rhode Island, Hospital, Providence, RI 02903, USA i COBRE Center for Cancer Research Development, Rhode Island Hospital, Providence, RI 02903, USA

POVEZANOST RADA