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Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology

Background Protein microarrays have enormous potential as in vitro diagnostic tools stemming from the ability to miniaturize whilst generating maximum evaluation of diagnostically relevant information from minute amounts of sample. In this report, we present a method known as repeatable arrays of p...

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Main Authors: Ashaari, Nur Suhanawati, Ramarad, Suganti, Khairuddin, Dzulaikha, Mat Akhir, Nor Azurah, Yuka, Hara, Mahadi, Nor Muhammad, Rahmah, Mohamed, Nathan, Sheila
Format: Article
Language:English
Published: BioMed Central 2015
Subjects:
TP Chemical technology
Online Access:http://eprints.intimal.edu.my/16/1/Development%20of%20Repeatable%20Arrays%20of%20Proteins%20using%20Immobilized%20DNA%20Microplate%20%28RAPID-M%29%20Technology.pdf
http://eprints.intimal.edu.my/16/
http://www.biomedcentral.com/
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spelling my-inti-eprints.162017-03-03T01:49:03Z http://eprints.intimal.edu.my/16/ Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology Ashaari, Nur Suhanawati Ramarad, Suganti Khairuddin, Dzulaikha Mat Akhir, Nor Azurah Yuka, Hara Mahadi, Nor Muhammad Rahmah, Mohamed Nathan, Sheila TP Chemical technology Background Protein microarrays have enormous potential as in vitro diagnostic tools stemming from the ability to miniaturize whilst generating maximum evaluation of diagnostically relevant information from minute amounts of sample. In this report, we present a method known as repeatable arrays of proteins using immobilized DNA microplates (RAPID-M) for high-throughput in situ protein microarray fabrication. The RAPID-M technology comprises of cell-free expression using immobilized DNA templates and in situ protein purification onto standard microarray slides. Results To demonstrate proof-of-concept, the repeatable protein arrays developed using our RAPID-M technology utilized green fluorescent protein (GFP) and a bacterial outer membrane protein (OmpA) as the proteins of interest for microarray fabrication. Cell-free expression of OmpA and GFP proteins using beads-immobilized DNA yielded protein bands with the expected molecular sizes of 27 and 30 kDa, respectively. We demonstrate that the beads-immobilized DNA remained stable for at least four cycles of cell-free expression. The OmpA and GFP proteins were still functional after in situ purification on the Ni–NTA microarray slide. Conclusion The RAPID-M platform for protein microarray fabrication of two different representative proteins was successfully developed. BioMed Central 2015 Article PeerReviewed text en http://eprints.intimal.edu.my/16/1/Development%20of%20Repeatable%20Arrays%20of%20Proteins%20using%20Immobilized%20DNA%20Microplate%20%28RAPID-M%29%20Technology.pdf Ashaari, Nur Suhanawati and Ramarad, Suganti and Khairuddin, Dzulaikha and Mat Akhir, Nor Azurah and Yuka, Hara and Mahadi, Nor Muhammad and Rahmah, Mohamed and Nathan, Sheila (2015) Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology. BMC Research Notes, 8 (1). ISSN 1756-0500 http://www.biomedcentral.com/ 10.1186/s13104-015-1637-3
institution INTI International University
building INTI Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider INTI International University
content_source INTI Institutional Repository
url_provider http://eprints.intimal.edu.my
language English
topic TP Chemical technology
spellingShingle TP Chemical technology
Ashaari, Nur Suhanawati
Ramarad, Suganti
Khairuddin, Dzulaikha
Mat Akhir, Nor Azurah
Yuka, Hara
Mahadi, Nor Muhammad
Rahmah, Mohamed
Nathan, Sheila
Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology
description Background Protein microarrays have enormous potential as in vitro diagnostic tools stemming from the ability to miniaturize whilst generating maximum evaluation of diagnostically relevant information from minute amounts of sample. In this report, we present a method known as repeatable arrays of proteins using immobilized DNA microplates (RAPID-M) for high-throughput in situ protein microarray fabrication. The RAPID-M technology comprises of cell-free expression using immobilized DNA templates and in situ protein purification onto standard microarray slides. Results To demonstrate proof-of-concept, the repeatable protein arrays developed using our RAPID-M technology utilized green fluorescent protein (GFP) and a bacterial outer membrane protein (OmpA) as the proteins of interest for microarray fabrication. Cell-free expression of OmpA and GFP proteins using beads-immobilized DNA yielded protein bands with the expected molecular sizes of 27 and 30 kDa, respectively. We demonstrate that the beads-immobilized DNA remained stable for at least four cycles of cell-free expression. The OmpA and GFP proteins were still functional after in situ purification on the Ni–NTA microarray slide. Conclusion The RAPID-M platform for protein microarray fabrication of two different representative proteins was successfully developed.
format Article
author Ashaari, Nur Suhanawati
Ramarad, Suganti
Khairuddin, Dzulaikha
Mat Akhir, Nor Azurah
Yuka, Hara
Mahadi, Nor Muhammad
Rahmah, Mohamed
Nathan, Sheila
author_facet Ashaari, Nur Suhanawati
Ramarad, Suganti
Khairuddin, Dzulaikha
Mat Akhir, Nor Azurah
Yuka, Hara
Mahadi, Nor Muhammad
Rahmah, Mohamed
Nathan, Sheila
author_sort Ashaari, Nur Suhanawati
title Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology
title_short Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology
title_full Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology
title_fullStr Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology
title_full_unstemmed Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology
title_sort development of repeatable arrays of proteins using immobilized dna microplate (rapid-m) technology
publisher BioMed Central
publishDate 2015
url http://eprints.intimal.edu.my/16/1/Development%20of%20Repeatable%20Arrays%20of%20Proteins%20using%20Immobilized%20DNA%20Microplate%20%28RAPID-M%29%20Technology.pdf
http://eprints.intimal.edu.my/16/
http://www.biomedcentral.com/
_version_ 1644541100546850816
score 13.211869

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