Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Affinity chromatography of plasma proteins (guanidinobenzoatase): use of mimetic matrices and mimetic soluble ligands to prevent the binding of albumin on target affinity matrices
Autore:
Murza, A; Aguilar, AR; Fernandez-Lafuente, R; Guisan, JM;
Indirizzi:
CSIC, Inst Catalisis, Dept Biocatalisis, Madrid 28049, Spain CSIC MadridSpain 28049 atalisis, Dept Biocatalisis, Madrid 28049, Spain Univ Complutense Madrid, Hosp Univ San Carlos, Dept Dermatol, E-28040 Madrid, Spain Univ Complutense Madrid Madrid Spain E-28040 atol, E-28040 Madrid, Spain
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 1, volume: 732, anno: 1999,
pagine: 165 - 172
SICI:
1387-2273(19990910)732:1<165:ACOPP
Fonte:
ISI
Lingua:
ENG
Soggetto:
SERUM-ALBUMIN; PURIFICATION; ASTROCYTES; CALCIUM;
Keywords:
mimetic matrices; mimetic ligands; enzymes; guanidinobenzoatase; albumin;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
18
Recensione:
Indirizzi per estratti:
Indirizzo: Guisan, JM CSIC, Inst Catalisis, Dept Biocatalisis, Campus UAM Cantoblanco, Madrid 28049, Spain CSIC Campus UAM Cantoblanco Madrid Spain 28049 id 28049, Spain
Citazione:
A. Murza et al., "Affinity chromatography of plasma proteins (guanidinobenzoatase): use of mimetic matrices and mimetic soluble ligands to prevent the binding of albumin on target affinity matrices", J CHROMAT B, 732(1), 1999, pp. 165-172

Abstract

Serum albumin is the most abundant protein in plasma and it has a high capacity to bind many small compounds and macromolecules. In this way, albuminmay promote important interferences during affinity chromatography of plasma proteins. Guanidinobenzoatase (GB) is a very relevant plasma protease that seems to be related to tumoral processes. This enzyme may be adsorbed ontailor-made agmatine-amide-agarose (CH-A) supports (e.g., the ones having 2 mu mol of guanidino groups per mi of agarose attached to the support, through a 6 C aliphatic chain). Such tailor-made supports containing a very low concentration of ionized groups are hardly able to adsorb any protein by anion-exchange. However, they are able to strongly adsorb albumin. In orderto solve this problem new mimetic affinity matrices have been designed: (i) by using the same ligand immobilized through a different chemical linkage[guanidino groups attached via secondary amino bonds, (AEA)] or (ii) by using slightly different Ligands (e.g., 1,8-octanediamine containing a primary amino group instead of a guanidino one) also attached to the support via amido bonds (CH-DAO). Albumin adsorbs on the target and on the two mimetic matrices while GB is mainly adsorbed on the target one. Moreover, the adsorption of albumin on the affinity matrix (CH-A) is very strongly inhibited by the presence of low concentrations of soluble ligands (e.g., 1,8-octanediamine containing two ionized primary amino groups). On the contrary, the adsorption of GB on CH-A is hardly inhibited by the presence of such mimetic soluble ligand. In this way, the former offering of crude GB samples to AEAplus the use of mimetic inhibitors during adsorption of the extract on CH-A completely prevent the undesirable adsorption of albumin. In a such way, an extremely selective adsorption of GB can be performed. Such an improved chromatography procedure allows a very easy affinity purification and detection of GB. (C) 1999 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/07/20 alle ore 05:24:02