R. Moerman et al., Miniaturized electrospraying as a technique for the production of microarrays of reproducible micrometer-sized protein spots, ANALYT CHEM, 73(10), 2001, pp. 2183-2189
Electrospraying in a stable cone-jet mode; at < 400 mum above a substrate i
s shown to be a powerful technique to produce arrays of identical micromete
r-sized spots consisting of biologically active substances. Aqueous solutio
ns with a surface tension of 0.04 N m(-1) and conductivities ranging from 0
.04 to 2.2 S m-l were sprayed at ultralow now rates ranging from 100 to 300
pL s(-1). The charged jet that emanates from the cone tip breaks up into a
spray of charged droplets that are deposited in the form of a uniform spot
of 130-350 mum in diameter by spraying during 0.5-3 s at 220-400 mum above
a substrate, respectively. After a spot was deposited, spraying was stoppe
d instantaneously by increasing the distance between the capillary-tip and
the substrate by an additional 100 mum using a computer-controlled x-y-z ta
ble. This was immediately followed by a rapid shift of the substrate 400 mu
m sideways and 100 mum upward, thus causing spraying to resume instantaneou
sly because of the increased electric field strength, which resulted in the
deposition of the next spot. It is shown here that spraying of lactate deh
ydrogenase (LDH), glucose-6-phosphate dehydrogenase (G6P-DH), and pyruvate
kinase (PK) on a liquid layer resulted in the complete preservation of thei
r activities despite the high solution conductivity of 3.3 S m-l and high c
urrents ranging from 300 to 500 nA. LDH and PK activities were fully preser
ved after spraying onto dry aluminum by adding 0.05 M buffer and 0.5 and 1
wt % of trehalose, respectively, to the spray solutions. Electrospraying al
lows for accurate dispensing of liquid volumes as small as 50 pL. Enzymatic
activities of LDH and PK are fully preserved after spraying.