title: Development of repeatable arrays of proteins using immobilized DNA microplate (RAPID-M) technology.
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.
  
  
  CONCLUSIONS
  The RAPID-M platform for protein microarray fabrication of two different representative proteins was successfully developed.
authors.affiliation: Malaysia Genome Institute, 43000, Bangi, Selangor DE, Malaysia. suhana_bio@yahoo.com.|Xynergen Sdn. Bhd., UKM Technology Centre, 43600, Bangi, Selangor DE, Malaysia. suhana_bio@yahoo.com.
authors.forename: Nur Suhanawati
authors.initials: NS
authors.lastname: Ashaari
authors.affiliation: Xynergen Sdn. Bhd., UKM Technology Centre, 43600, Bangi, Selangor DE, Malaysia. suga4u2c@gmail.com.
authors.forename: Suganti
authors.initials: S
authors.lastname: Ramarad
authors.affiliation: Malaysia Genome Institute, 43000, Bangi, Selangor DE, Malaysia. dzulaikha.k@gmail.com.|Xynergen Sdn. Bhd., UKM Technology Centre, 43600, Bangi, Selangor DE, Malaysia. dzulaikha.k@gmail.com.
authors.forename: Dzulaikha
authors.initials: D
authors.lastname: Khairuddin
authors.affiliation: Malaysia Genome Institute, 43000, Bangi, Selangor DE, Malaysia. azurah@mfrlab.org.|Xynergen Sdn. Bhd., UKM Technology Centre, 43600, Bangi, Selangor DE, Malaysia. azurah@mfrlab.org.
authors.forename: Nor Azurah Mat
authors.initials: NA
authors.lastname: Akhir
authors.affiliation: School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor DE, Malaysia. yuka.hara@newinti.edu.my.|INTI International University, Bandar Baru Nilai, 71800, Nilai, Negeri Sembilan, Malaysia. yuka.hara@newinti.edu.my.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Hara
authors.affiliation: Malaysia Genome Institute, 43000, Bangi, Selangor DE, Malaysia. nor.mahadi@gmail.com.
authors.forename: Nor Muhammad
authors.initials: NM
authors.lastname: Mahadi
authors.affiliation: School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor DE, Malaysia. rahmah.mohamed@newinti.edu.my.|INTI International University, Bandar Baru Nilai, 71800, Nilai, Negeri Sembilan, Malaysia. rahmah.mohamed@newinti.edu.my.
authors.forename: Rahmah
authors.initials: R
authors.lastname: Mohamed
authors.affiliation: Malaysia Genome Institute, 43000, Bangi, Selangor DE, Malaysia. sheila@ukm.edu.my.|School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor DE, Malaysia. sheila@ukm.edu.my.
authors.forename: Sheila
authors.initials: S
authors.lastname: Nathan
mesh_terms: D001425:Bacterial Outer Membrane Proteins; D002474:Cell-Free System; D049452:Green Fluorescent Proteins; D061925:Immobilized Nucleic Acids; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D061925:Immobilized Nucleic Acids; D049452:Green Fluorescent Proteins; C078167:OMPA outer membrane proteins