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Titolo:
AFFINITY PURIFICATION OF IMMUNOGLOBULIN-M USING A NOVEL SYNTHETIC LIGAND
Autore:
PALOMBO G; VERDOLIVA A; FASSINA G;
Indirizzi:
TECNOGEN SCPA,PARCO SCI I-81015 PIANA MONTE VERNA CE ITALY TECNOGEN SCPA I-81015 PIANA MONTE VERNA CE ITALY
Titolo Testata:
Journal of chromatography B. Biomedical sciences and applications
fascicolo: 1, volume: 715, anno: 1998,
pagine: 137 - 145
SICI:
0378-4347(1998)715:1<137:APOIUA>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
MANNAN-BINDING-PROTEIN; STEP PURIFICATION; CHROMATOGRAPHY; ANTIBODIES;
Keywords:
IMMUNOGLOBULINS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
18
Recensione:
Indirizzi per estratti:
Citazione:
G. Palombo et al., "AFFINITY PURIFICATION OF IMMUNOGLOBULIN-M USING A NOVEL SYNTHETIC LIGAND", Journal of chromatography B. Biomedical sciences and applications, 715(1), 1998, pp. 137-145

Abstract

While monoclonal antibodies of the G class can be conveniently purified by affinity chromatography using immobilized protein A or G, even on a large scale, scaling up IgM purification still presents several problems, since specific and cost-effective ligands for IgM are not available. A synthetic peptide (TG19318), deduced from the screening of a combinatorial peptide library, was characterized previously by our group for its binding properties for immunoglobulins of the G class and its applicability as a synthetic ligand for polyclonal and monoclonal IgG purification, from sera or cell culture supernatants. In this study, we have examined the ligand recognition properties for IgM, immobilizing the synthetic peptide on different affinity supports and examining its ability to purify IgMs from serum, ascitic fluid and cell culture supernatants. TG19318 affinity columns proved useful for a very convenient one-step purification of monoclonal IgMs directly from crude sources, loading the samples on the columns equilibrated with saline buffers at pH values ranging from 5 to 7, and eluting adsorbed IgM by a buffer change to 0.1 M acetic acid or 0.05-0.1 M sodium bicarbonate, pH9.0. Antibody purity after affinity purification was very high, closeto 85-95%, as determined by densitometric scanning of sodium dodecyl sulfate-polyacrylamide gels of purified fractions, and by gel permeation analysis. Antibody activity was fully recovered after purification,as determined by immunoassays. Column capacity was related to the type of support used for ligand immobilization, and ranged from 2 to 8 mgof IgM/ml of support. (C) 1998 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 08/04/20 alle ore 12:12:04