Micropatterned agarose gels for stamping arrays of proteins and gradients of proteins

Proteomics. 2004 Aug;4(8):2366-76. doi: 10.1002/pmic.200300748.

Abstract

We describe a method for repetitive and rapid formation of planar microarrays and gradients of proteins using patterned agarose stamps. It demonstrates: (i) micropatterning of agarose gels with feature sizes as small as 2 microm; (ii) inking of posts (diameter 50-1000 microm) on patterned agarose stamps with one or multiple (here, eight) proteins and repetitive stamping of patterns (>100 times in the case of one protein) and arrays (20 times in the case of eight proteins) without the need for intermediate re-inking; (iii) transferring spots of proteins with good homogeneity in surface coverage to glass slides; (iv) applying this technique to surface-based immunoassays; (v) stamping that requires only sub-nanomolar amounts of protein (typically approximately 3 microg in approximately 0.6 microL of solution); (vi) stamping without the need for drying of the proteins, as opposed to stamping with stamps made of poly(dimethylsiloxane); and (vii) patterning gradients of proteins by allowing two proteins to diffuse toward each other in an agarose stamp, followed by printing the protein gradients onto a surface.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Gels / chemistry*
  • Immunoassay* / instrumentation
  • Immunoassay* / methods
  • Ink
  • Microscopy, Fluorescence
  • Protein Array Analysis / instrumentation
  • Protein Array Analysis / methods
  • Proteins / analysis*
  • Sepharose / chemistry*
  • Surface Properties

Substances

  • Gels
  • Proteins
  • Sepharose