Rp. Huang, Simultaneous detection of multiple proteins with an array-based enzyme-linked immunosorbent assay (ELISA) and enhanced chemiluminescence (ECL), CLIN CH L M, 39(3), 2001, pp. 209-214
Protein arrays hold a promise in basic and clinical applications. As the fi
rst step to develop such array system, I used an array-based enzyme-linked
immunosorbent assay (ELISA) and enhanced chemiluminescence (ECL) to demonst
rate the feasibility of simultaneous detection of multiple proteins. In the
direct ELISA system, different known immunoglobulin Gs (IgGs) were immobil
ized onto polyvinylidine difloride (PDVF) membrane through 96-well format B
io-Dot unit. The antigens were then individually and collectively detected
by incubation of membranes with different antibodies coupled with ECL. In t
he sandwich ELISA system, the cytokine capture antibodies were immobilized
onto PDVF membranes. The membranes were then incubated with single cytokine
or a combination of different cytokines. The captured cytokines were detec
ted by biotin-conjugated antibodies coupled with ECL system. Experiments de
monstrated that multiple IgGs and cytokines could be simultaneously detecte
d using this approach with high specificity and sensitivity. More important
ly, cytokines from biological samples were detected using this approach, wh
ich can be used in any general laboratory setting without any sophisticated
equipment. This concept could be extended to develop a protein-based array
system.