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Dot-ELISA Affinity Test: An Easy, Low-Cost Method to Estimate Binding Activity of Monoclonal Antibodies | OMICS International | Abstract
ISSN: 2155-9872

Journal of Analytical & Bioanalytical Techniques
Open Access

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Research Article

Dot-ELISA Affinity Test: An Easy, Low-Cost Method to Estimate Binding Activity of Monoclonal Antibodies

Svobodova Z1*, Jankovicova B1 , Horak D2 and Bilkova Z1

1Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, 532 10 Pardubice, Czech Republic

2Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovsky Sq. 2, 162 06 Prague 6, Czech Republic

*Corresponding Author:
Svobodova Zuzana
Department of Biological and Biochemical Sciences
Faculty of Chemical Technology, University of Pardubice
Studentska 573, 532 10 Pardubice, Czech Republic
Tel: +420 46 6037700

Received date: June 07, 2013; Accepted date: June 26, 2013; Published date: June 28, 2013

Citation: Svobodova Z, Jankovicova B, Horak D, Bilkova Z (2013) Dot-ELISA Affinity Test: An Easy, Low-Cost Method to Estimate Binding Activity of Monoclonal Antibodies. J Anal Bioanal Tech 4:168. doi: 10.4172/2155-9872.1000168

Copyright: © 2013 Svobodova Z, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Selecting the “right” monoclonal antibody (mAb) for an immunoaffinity-based application can be tricky, as many mAb producers offer a wide range of mAb clones against molecular structures of interest. Since there are significant differences in the quality of mAb clones, and particularly in their binding activity, an easy method for quick and low-cost comparison of various mAb clones was developed. The dot-ELISA affinity test is a simple, versatile and instrumentally no demanding technique, since it requires no expensive equipment (such as an ELISA reader or chemiluminescence/fluorescence imaging system) and can be performed in any biochemical laboratory. This method is based on a previously described dot-ELISA technique that is improved with a chaotropic step using different concentrations of ammonium thiocyanate in the range 0-2 M. In this work, the dot-ELISA affinity test was optimized on Aβ peptide as antigen and anti-Aβ mAb. Such protocol was then applied to a panel of eight anti-EpCAM (epithelial cell adhesion molecule) mAbs which should be subsequently used for preparation of magnetic immunosorbent to capture circulating tumor cells (CTCs).