Engineering resonance energy transfer for advanced immunoassays: The case of celiac disease

Sabato D'Auria, Elisa Apicella, Maria Staiano, Stefano Di Giovanni, Giuseppe Ruggiero, Mauro Rossi, Pabak Sarkar, Rafal Luchowski, Ignacy Gryczynski, Zygmunt Gryczynski

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Celiac disease (CD) is an immune-mediated disorder affecting genetically predisposed subjects. It is caused by the ingestion of wheat gluten and related prolamins. A final diagnosis for this disease can be obtained by examination of jejunal biopsies. Nevertheless, different analytical approaches have been established to detect the presence of anti-tissue transglutaminase antibodies that represent a serological hallmark of the disease. In this work, we explored a new method for the diagnosis of CD based on the detection of serum anti-transglutaminase antibodies by resonance energy transfer (RET) between donor molecules and acceptor molecules. In particular, we labeled the liver transglutaminase (tTG) enzyme from guinea pig and the rabbit anti-tTG antibodies with a couple of fluorescence probes that are able to make RET if they are located within with Förster distance. We labeled tTG with the fluorescence probe DyLight 594 as donor and the anti-tTG antibodies with the fluorescence probe DyLight 649 as acceptor. However, due to the large size of the formed complex (tTG/anti-tTG), and consequently to the low efficiency energy transfer process between the donor-acceptor molecules, we explored a new experimental approach that allows us to extend the utilizable range of RET between donor:acceptor pairs by using one single molecule as donor and multiple molecules as energy acceptors, instead of using a single acceptor molecule as usually occurs in RET experiments. The obtained results clearly show that the use of one donor and multiacceptor strategy enables for a simple and rapid detection of serum anti-transglutaminase antibodies. In addition, our results point out that it is possible to consider this approach as a new method for a wide variety of analytical assays.

Original languageEnglish
Pages (from-to)13-17
Number of pages5
JournalAnalytical Biochemistry
Volume425
Issue number1
DOIs
StatePublished - 1 Jun 2012

Fingerprint

Energy Transfer
Celiac Disease
Immunoassay
Energy transfer
Transglutaminases
Anti-Idiotypic Antibodies
Molecules
Fluorescence
Prolamins
Antibodies
Glutens
Immune System Diseases
Serum
Biopsy
Triticum
Guinea Pigs
Liver
Eating
Assays
Rabbits

Keywords

  • Fluorescence
  • Proteins
  • Resonance energy transfer

Cite this

D'Auria, Sabato ; Apicella, Elisa ; Staiano, Maria ; Di Giovanni, Stefano ; Ruggiero, Giuseppe ; Rossi, Mauro ; Sarkar, Pabak ; Luchowski, Rafal ; Gryczynski, Ignacy ; Gryczynski, Zygmunt. / Engineering resonance energy transfer for advanced immunoassays : The case of celiac disease. In: Analytical Biochemistry. 2012 ; Vol. 425, No. 1. pp. 13-17.
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abstract = "Celiac disease (CD) is an immune-mediated disorder affecting genetically predisposed subjects. It is caused by the ingestion of wheat gluten and related prolamins. A final diagnosis for this disease can be obtained by examination of jejunal biopsies. Nevertheless, different analytical approaches have been established to detect the presence of anti-tissue transglutaminase antibodies that represent a serological hallmark of the disease. In this work, we explored a new method for the diagnosis of CD based on the detection of serum anti-transglutaminase antibodies by resonance energy transfer (RET) between donor molecules and acceptor molecules. In particular, we labeled the liver transglutaminase (tTG) enzyme from guinea pig and the rabbit anti-tTG antibodies with a couple of fluorescence probes that are able to make RET if they are located within with F{\"o}rster distance. We labeled tTG with the fluorescence probe DyLight 594 as donor and the anti-tTG antibodies with the fluorescence probe DyLight 649 as acceptor. However, due to the large size of the formed complex (tTG/anti-tTG), and consequently to the low efficiency energy transfer process between the donor-acceptor molecules, we explored a new experimental approach that allows us to extend the utilizable range of RET between donor:acceptor pairs by using one single molecule as donor and multiple molecules as energy acceptors, instead of using a single acceptor molecule as usually occurs in RET experiments. The obtained results clearly show that the use of one donor and multiacceptor strategy enables for a simple and rapid detection of serum anti-transglutaminase antibodies. In addition, our results point out that it is possible to consider this approach as a new method for a wide variety of analytical assays.",
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D'Auria, S, Apicella, E, Staiano, M, Di Giovanni, S, Ruggiero, G, Rossi, M, Sarkar, P, Luchowski, R, Gryczynski, I & Gryczynski, Z 2012, 'Engineering resonance energy transfer for advanced immunoassays: The case of celiac disease', Analytical Biochemistry, vol. 425, no. 1, pp. 13-17. https://doi.org/10.1016/j.ab.2012.02.035

Engineering resonance energy transfer for advanced immunoassays : The case of celiac disease. / D'Auria, Sabato; Apicella, Elisa; Staiano, Maria; Di Giovanni, Stefano; Ruggiero, Giuseppe; Rossi, Mauro; Sarkar, Pabak; Luchowski, Rafal; Gryczynski, Ignacy; Gryczynski, Zygmunt.

In: Analytical Biochemistry, Vol. 425, No. 1, 01.06.2012, p. 13-17.

Research output: Contribution to journalArticle

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T2 - The case of celiac disease

AU - D'Auria, Sabato

AU - Apicella, Elisa

AU - Staiano, Maria

AU - Di Giovanni, Stefano

AU - Ruggiero, Giuseppe

AU - Rossi, Mauro

AU - Sarkar, Pabak

AU - Luchowski, Rafal

AU - Gryczynski, Ignacy

AU - Gryczynski, Zygmunt

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Y1 - 2012/6/1

N2 - Celiac disease (CD) is an immune-mediated disorder affecting genetically predisposed subjects. It is caused by the ingestion of wheat gluten and related prolamins. A final diagnosis for this disease can be obtained by examination of jejunal biopsies. Nevertheless, different analytical approaches have been established to detect the presence of anti-tissue transglutaminase antibodies that represent a serological hallmark of the disease. In this work, we explored a new method for the diagnosis of CD based on the detection of serum anti-transglutaminase antibodies by resonance energy transfer (RET) between donor molecules and acceptor molecules. In particular, we labeled the liver transglutaminase (tTG) enzyme from guinea pig and the rabbit anti-tTG antibodies with a couple of fluorescence probes that are able to make RET if they are located within with Förster distance. We labeled tTG with the fluorescence probe DyLight 594 as donor and the anti-tTG antibodies with the fluorescence probe DyLight 649 as acceptor. However, due to the large size of the formed complex (tTG/anti-tTG), and consequently to the low efficiency energy transfer process between the donor-acceptor molecules, we explored a new experimental approach that allows us to extend the utilizable range of RET between donor:acceptor pairs by using one single molecule as donor and multiple molecules as energy acceptors, instead of using a single acceptor molecule as usually occurs in RET experiments. The obtained results clearly show that the use of one donor and multiacceptor strategy enables for a simple and rapid detection of serum anti-transglutaminase antibodies. In addition, our results point out that it is possible to consider this approach as a new method for a wide variety of analytical assays.

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D'Auria S, Apicella E, Staiano M, Di Giovanni S, Ruggiero G, Rossi M et al. Engineering resonance energy transfer for advanced immunoassays: The case of celiac disease. Analytical Biochemistry. 2012 Jun 1;425(1):13-17. https://doi.org/10.1016/j.ab.2012.02.035