Naslov
Use of proteolytic enzymes immobilized on monolithic CIM disks for rapid analytical and preparative cleavage of proteins

Autori
Y.P. Lim, M. Ručević, H. Callanan, M.K. Brown, D.C. Hixson, D. Josić.

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
24th International Symphosium on the Separation of Proteins, Peptides and Polynucleotides

Mjesto i datum
Aachen, Njemačka, 19.10.2004. - 22.10.2004

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
Convective Interaction Media; monolithic disks; proteolythic enzymes

Sažetak
So called Convective Interaction Media (CIM) monolithic disks offer a simple and rapid method for proteolythic cleavage of investigated proteins. We used several proteolytic enzymes such as trypsin, elastase and granzyme immobilized on CIM disks of various sizes for digestion of standard proteins and different samples with unknown proteins. Proteolytic fragments can be subsequently separated by use of SDS-PAGE or RP-HPLC. For RP-HPLC particle-based or disk-shaped chromatographic columns can be used. Unknown proteins can be identified by Western blot, SELDI-TOF technique or by use of mass spectrometry. The immobilized enzymes on the Connective Interaction Media are stable and still active after repeated runs. A partial or complete digestion can be achieved by varying the flow rate.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biotehnologija

POVEZANOST RADA