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Protein Microarray Delivery Methods

Protein and Antibody Microarrays

Creation and Manufacture of Protein Chips

 

Protein Delivery Methods
            The manufacture of microarrays is a highly automated process which involves imprinting of capture molecules on a bio-reactive film on an array or slide surface in a 2D array format (50).  The spacing of the spots depends on the size of the capture agents.  Antibody arrays typically have a 375 mm spacing (31).  Peptides may be synthesized on planar surfaces using photolithography (51,52) or SPOT technology (53).  Photolithography is known to be the first method adapted to be used with protein arrays (52). Zyomyx has constructed silicon arrays with photolithography capable of detecting more than 10,000 proteins (9).
            96-dot blot instruments have been used previously to create low-density protein arrays on filters (54,22), however high-density protein microarrays with greater than 30 000 spots per slide may be achieved using robotic contact printing instruments, similar to those for DNA microarrays (31,26).  Sub-nanoliter sample volumes are delivered directly to the surface of chips using tiny pins of contact printing arrayers.  However, due to the fact that contact printing robots are unable to align their pins to pre-fabricated structures and need to touch the surface, non-contact robotic printers similar to in-jet technology were used to deposit nanoliter to picoliter protein droplets to polyacrylamide gel packets (55) and nanowells (36).  The shearing force during drop formation of the current Packard ink-jet microarrayers is thought to damage some samples (56).  More recently, electrospray deposition technology was applied to deliver dry proteins to a dextran-grafted surface (57).  This technology reduced the spot size from ~150 mm to 30 mm.

Next: Protein Chip Capture Molecules and Their Limitations

 

References for Protein and Antibody Microarrays

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Introduction and Background to Protein Chips and Antibody Chips.

Types of Antibody and Protein Chips

 

 

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