Micropatterned agarose gels for stamping arrays of proteins and gradients of proteins
Micropatterned agarose gels for stamping arrays of proteins and gradients of proteins
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DOI：
10.1002/pmic.200300748
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发表时间：
2004-08-01
期刊：
影响因子：
3.4
通讯作者：
Whitesides, GM
中科院分区：
文献类型：
--
作者：
Mayer, M;Yang, J;Whitesides, GM
我们描述了一种使用图案化琼脂糖印章重复和快速形成平面微阵列和蛋白质梯度的方法。它演示了：(i) 特征尺寸小至 2 微米的琼脂糖凝胶微图案化； (ii) 用一种或多种（此处为八种）蛋白质在带图案的琼脂糖印章上给柱（直径 50-1000 微米）着墨，并重复印制图案（在一种蛋白质的情况下 >100 次）和阵列（在八种蛋白质的情况下 20 次），无需中间重新上墨； (iii) 将表面覆盖均匀性良好的蛋白质斑点转移到载玻片上； (iv) 将该技术应用于基于表面的免疫测定； (v) 冲压仅需要亚纳摩尔量的蛋白质（通常为 g0.6 杯溶液中的 -3 杯）； (vi) 不需要干燥蛋白质的冲压，与用聚(二甲基硅氧烷)制成的印模冲压不同； (vii) 通过允许两种蛋白质在琼脂糖印章中相互扩散来形成蛋白质梯度图案，然后将蛋白质梯度打印到表面上。
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 mum; (ii) inking of posts (diameter 50-1000 mum) 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 -3 mug in g0.6 mug 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.