Generation and characterization of D-dimer specific monoclonal antibodies for use in latex agglutination test

Beáta Török-Nagy, József Antal, B. Dénes

Research output: Contribution to journalArticle

Abstract

The commercially available D-dimer assays used in the clinical practice often show differences in the results, and their specificity and sensitivity are rather unsatisfactory. Our aim was to develop a new monoclonal antibody against D-dimer with a proper specificity, and estimating its suitability using in a latex agglutination diagnostic test. Monoclonal antibodies were generated using hybridoma technology. Their titer was determined by a self-developed ELISA method. The cross-reactions of the antibodies were tested. Characterization of the epitope specificity of a selected antibody was performed through digestion of D-dimer followed by Western blotting. The amino acid sequences of the active antigen fragments were determined. According to the ELISA results, 38 cell groups were constated as antibody-producing hybridomas, among them 7 gave raised titer of antibody and were cloned. Based on the cross-reaction analysis, none of the antibodies gave cross-reaction with fibrin-E and fibrinogen-E fragments but reacted with fibrin D and fibrinogen D fragments. A low cross-reaction was showed with fibrinogen and fibrin X and Y. Contrary to the others, antibody 2B9 gave no cross-reaction with fibrinogen and reacted weakly with fibrin X and Y fragments. According to the epitope analysis the antibody 2B9 binds to amino acids 94–99 and to amino acids 140–147 on the beta chain and it recognizes the amino acids 23–32 and 93–98 on the gamma chain of D-dimer. Considering the characteristics of the above mentioned monoclonal antibody 2B9, we found that it is suitable to be a basis for a D-dimer diagnostic test with proper specificity.

Original languageEnglish
Article numbere0212104
JournalPloS one
Volume14
Issue number2
DOIs
Publication statusPublished - Feb 1 2019

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Latex Fixation Tests
latex agglutination test
Latex
monoclonal antibodies
cross reaction
Cross Reactions
fibrin
Monoclonal Antibodies
fibrinogen
antibodies
Antibodies
Fibrin
hybridomas
Amino Acids
Hybridomas
diagnostic techniques
epitopes
amino acids
Routine Diagnostic Tests
Fibrinogen

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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Generation and characterization of D-dimer specific monoclonal antibodies for use in latex agglutination test. / Török-Nagy, Beáta; Antal, József; Dénes, B.

In: PloS one, Vol. 14, No. 2, e0212104, 01.02.2019.

Research output: Contribution to journalArticle

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