Development of repeatable arrays of proteins using immobilized DNA microplate (RAPID-M) technology Biotechnology

Nur Suhanawati Ashaari, Suganti Ramarad, Dzulaikha Khairuddin, Nor Azurah Mat Akhir, Yuka Hara, Nor Muhammad Mahadi, Rahmah Mohamed, Sheila Nathan

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Article number669
JournalBMC Research Notes
Volume8
Issue number1
DOIs
Publication statusPublished - 12 Nov 2015

Fingerprint

Immobilized Nucleic Acids
Protein Array Analysis
Biotechnology
Technology
Proteins
Microarrays
Green Fluorescent Proteins
Fabrication
Bacterial Outer Membrane Proteins
Purification
Cell Cycle
Throughput

Keywords

  • Cell-free expression
  • Microarray fabrication
  • Protein array
  • RAPID-M

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Development of repeatable arrays of proteins using immobilized DNA microplate (RAPID-M) technology Biotechnology. / Ashaari, Nur Suhanawati; Ramarad, Suganti; Khairuddin, Dzulaikha; Akhir, Nor Azurah Mat; Hara, Yuka; Mahadi, Nor Muhammad; Mohamed, Rahmah; Nathan, Sheila.

In: BMC Research Notes, Vol. 8, No. 1, 669, 12.11.2015.

Research output: Contribution to journalArticle

Ashaari, Nur Suhanawati ; Ramarad, Suganti ; Khairuddin, Dzulaikha ; Akhir, Nor Azurah Mat ; Hara, Yuka ; Mahadi, Nor Muhammad ; Mohamed, Rahmah ; Nathan, Sheila. / Development of repeatable arrays of proteins using immobilized DNA microplate (RAPID-M) technology Biotechnology. In: BMC Research Notes. 2015 ; Vol. 8, No. 1.
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