Biomedical Engineering Reference
In-Depth Information
(a)
Protein
Protein
Antibody
Lysate
(b)
P
P
Antibody
Kinase
(c)
H5N1
HA/Virus
H3N2
H1N1
FIGURE 13.6 Biological applications of SMMs: (a) the process of screening cellular lysates
without prior protein purification; (b) peptide microarray used for kinase substrate identifica-
tion; (c) carbohydrate microarray used for virus detection.
were observed capable of cleaving their corresponding substrates selectively. In a
similar setup, Salisbury et al. synthesized a combinatorial library with 361 peptide
coumarin conjugates and used it to study the substrate fingerprints of proteases on the
array [61]. The group was able to obtain a comprehensive proteolytic fingerprint with
a thrombin enzyme in a single experiment. The approach also provided important
quantitative data to determine the preference of the substrates, in this case kcat/km.
Gosalia and Diamond developed a novel nanodroplet SMM approach to circum-
vent the tedious immobilization process. In their study, a library of 352 compounds
without any tag was first dispensed in glycerol droplets onto a plain glass slide [62].
Following that, the enzyme and its corresponding substrate were added in aerosol
form to the discrete nanodroplets on the glass slide. The printed nanodroplets can
last for a considerably long period during the screening process because of the high
boiling point of glycerol. With this method, the group was able to obtain distinct
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