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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.


IMPACT FACTOR increased in 2015: 3.017
Rank 5 out of 30 in category Medical Laboratory Technology in the 2014 Thomson Reuters Journal Citation Report/Science Edition

SCImago Journal Rank (SJR) 2015: 0.873
Source Normalized Impact per Paper (SNIP) 2015: 0.982
Impact per Publication (IPP) 2015: 2.238

249,00 € / $374.00 / £187.00*

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ISSN
1437-4331
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Quality Control of Coated Antibodies: New, Rapid Determination of Binding Affinity

Rémy Ricoux / Bénédicte Chazaud / Jean-Pierre Tresca / Michel Pontet

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 38, Issue 3, Pages 239–243, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2000.035, June 2005

Publication History

Published Online:
2005-06-01

Abstract

A procedure is described for the determination of the affinity constant between a fluid-phase biotinylated antigen and a solid-phase monoclonal antibody. This procedure allows evaluation of the efficiency of an antibody as a coated tool for an immunoassay. For this purpose, the biotinylation of the antigen and its further quantitative measurement by streptavidin-peroxidase led to a single reversible interaction, the binding affinity of which greatly determines the quality of the assay. The free and bound fractions of the biotinylated antigen were obtained in wells coated with a low level of immobilized antibodies. At the equilibrium state, the free antigen present in the supernatant of these wells was further transferred to high level antibody coated wells which captured all the free antigen molecules. These molecules were quantified using a standard curve established with known concentrations of biotinylated antigen, also incubated in wells coated with the high level of antibody.

Citing Articles

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[1]
Nahla A. Hussein, Yasser E. Shahein, Amr E. El-Hakim, Amira M. Abouelella, Rasha A. Guneidy, and Ragaa R. Hamed
International Journal of Biological Macromolecules, 2014, Volume 67, Page 7
[2]
Yasser E. Shahein, Mohamed T. Abd El-Rahim, Nahla A. Hussein, Ragaa R. Hamed, Amr E. El-Hakim, and Maged M. Barakat
International Journal of Biological Macromolecules, 2010, Volume 47, Number 5, Page 614

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