Document Detail


Peroxidase-labelling of human serum transferrin by conjugation to oligosaccharide moieties.
MedLine Citation:
PMID:  9694587     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The generation of reactive aldehydes on the carbohydrate moieties of the human serum transferrin was performed by a derivatization procedure based on the mild oxidation with sodium periodate and subsequent reaction with peroxidase hydrazide. The synthesized conjugate was compared to that obtained by modification of the amino acid side chains of transferrin. The conjugate reaction mixture assayed by SDS-PAGE consisted, besides unreacted compounds, of three main bands, corresponding to a molar ratio transferrin:peroxidase of 1:1, 1:2, 1:3. After blotting, these bands were identified by either anti-peroxidase and anti-transferrin antibodies on nitrocellulose membrane. ELISA detection method showed that the conjugate via oligosaccharide moieties (glycans) was still recognized not only by the anti-transferrin antibodies but also by the specific cellular receptor, while the conjugate via amino acids failed to display this latter ability. The different behaviour can be probably due to a significant damage of the protein structure or, possibly, to the peroxidase binding at sites recognized by the receptor. The results reported here indicate that the conjugation procedure through glycans leads to stable and selected transferrin-conjugates fully exhibiting their biological activity.
Authors:
A M D'Alessandro; G D'Andrea; A Oratore
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Clinica chimica acta; international journal of clinical chemistry     Volume:  274     ISSN:  0009-8981     ISO Abbreviation:  Clin. Chim. Acta     Publication Date:  1998 Jun 
Date Detail:
Created Date:  1998-10-13     Completed Date:  1998-10-13     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  1302422     Medline TA:  Clin Chim Acta     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  189-97     Citation Subset:  IM    
Affiliation:
Department of Biomedical Sciences and Technologies, University of L'Aquila, Italy. dalessandro@aquila.infn.it
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MeSH Terms
Descriptor/Qualifier:
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Humans
Immunoenzyme Techniques*
Oligosaccharides*
Peroxidase / metabolism*
Receptors, Transferrin / metabolism
Transferrin / analysis*,  metabolism
Chemical
Reg. No./Substance:
0/Oligosaccharides; 0/Receptors, Transferrin; 11096-37-0/Transferrin; EC 1.11.1.7/Peroxidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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