Printing of protein microarrays via a capillary‐free fluid jetting mechanism

Current proteomics experiments rely upon printing techniques such as ink jet, pin, or quill arrayers that were developed for the creation of cDNA microarrays. These techniques often do not meet the requirements needed for successful spotting of proteins to perform high‐throughput, array‐based proteomic profiling. Biological laser printing (BioLP) is a spotting technology that does not rely on solid pins, quill pins, or capillary‐based fluidics. The non‐contact mechanism of BioLP utilizes a focused laser pulse to transfer protein solutions, thereby eliminating the potential for orifice clogging, air bubbles, and unnecessary volume loss potentially encountered in commercially available spotting technologies. The speed and spot‐to‐spot reproducibility of BioLP is comparable to other techniques, while the minimum spot diameter and volume per printed droplet is significantly less at 30 µm and ∼500 fL, respectively. The transfer of fluid by BioLP occurs through a fluid jetting mechanism, as observed by high‐speed images of the printing process. Arraying a solution of BSA with subsequent immunodetection demonstrates the reproducible spotting of protein in an array format with CVs of <3%. Printing of the enzyme alkaline phosphatase followed by a positive reaction with a colorimetric substrate demonstrates that functional protein can be spotted using this laser‐based printer.

[1]  Bradley R Ringeisen,et al.  Laser printing of pluripotent embryonal carcinoma cells. , 2004, Tissue engineering.

[2]  D. Krizman,et al.  Picoliter‐Scale Protein Microarrays by Laser Direct Write , 2002, Biotechnology progress.

[3]  Costas Fotakis,et al.  Shadowgraphic imaging of the sub-ps laser-induced forward transfer process , 2002 .

[4]  L. Liotta,et al.  Proteomic profiling of the cancer microenvironment by antibody arrays , 2001, Proteomics.

[5]  Hackjin Kim,et al.  Ultrafast Dynamics of Shock Waves in Polymers and Proteins: The Energy Landscape , 1999 .

[6]  B R Ringeisen,et al.  Generation of mesoscopic patterns of viable Escherichia coli by ambient laser transfer. , 2002, Biomaterials.

[7]  Virginia Espina,et al.  Protein microarrays: Molecular profiling technologies for clinical specimens , 2003, Proteomics.

[8]  J A Barron,et al.  Biological Laser Printing: A Novel Technique for Creating Heterogeneous 3-dimensional Cell Patterns , 2004, Biomedical microdevices.

[9]  S. Schreiber,et al.  Printing proteins as microarrays for high-throughput function determination. , 2000, Science.

[10]  B. Haab.  Methods and applications of antibody microarrays in cancer research , 2003, Proteomics.

[11]  M. Snyder,et al.  Analyzing antibody specificity with whole proteome microarrays , 2003, Nature Biotechnology.

[12]  Frances S Ligler,et al.  Method for printing functional protein microarrays. , 2003, BioTechniques.

[13]  William C Reinhold,et al.  Proteomic profiling of the NCI-60 cancer cell lines using new high-density reverse-phase lysate microarrays , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Alberto Piqué,et al.  Time-resolved optical microscopy of a laser-based forward transfer process , 2001 .

[15]  B R Ringeisen,et al.  Biological laser printing of genetically modified Escherichia coli for biosensor applications. , 2004, Biosensors & bioelectronics.

[16]  Barry J. Spargo,et al.  Biological laser printing of three dimensional cellular structures , 2004 .

[17]  Dieter Stoll,et al.  Protein microarrays: Promising tools for proteomic research , 2003, Proteomics.

[18]  R. A. McGill,et al.  Laser transfer of biomaterials: Matrix-assisted pulsed laser evaporation (MAPLE) and MAPLE Direct Write , 2003 .

[19]  S M Hanash,et al.  Protein based microarrays: A tool for probing the proteome of cancer cells and tissues , 2001, Proteomics.

[20]  P. Angenendt,et al.  Protein and antibody microarray technology. , 2003, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[21]  E. Petricoin,et al.  Reverse phase protein microarrays which capture disease progression show activation of pro-survival pathways at the cancer invasion front , 2001, Oncogene.

[22]  R. A. McGill,et al.  Laser deposition of polymer and biomaterial films. , 2003, Chemical reviews.