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

BMC Research Notes

Sheila Nathan⁴  Rahmah Mohamed³  Nor Muhammad Mahadi¹  Yuka Hara³  Nor Azurah Mat Akhir²  Dzulaikha Khairuddin²  Suganti Ramarad²  Nur Suhanawati Ashaari² 

[1] Malaysia Genome Institute, Bangi, 43000, Selangor DE, Malaysia;Xynergen Sdn. Bhd., UKM Technology Centre, Bangi, 43600, Selangor DE, Malaysia;INTI International University, Bandar Baru Nilai, Nilai, 71800, Negeri Sembilan, Malaysia;School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, 43600, Selangor DE, Malaysia

关键词: RAPID-M;    Protein array;    Microarray fabrication;    Cell-free expression;   

Others  :  1232677
DOI  :  10.1186/s13104-015-1637-3

 received in 2015-04-20, accepted in 2015-10-26,  发布年份 2015

【 摘 要 】

Background

Protein microarrays have enormouspotential 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.

【 授权许可】

   
2015 Ashaari et al.

【 图 表 】

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