ELISA incubation times can be reduced by 2.45-GHz microwaves.
The ELISA (Enzyme Linked Immuno Sorbent Assay) technique is widely applied in the field of immunology. The use of this technique implies several incubation periods, often requiring more than one hour each. Microwave irradiation is known to be very useful in accelerating different kinds of processes. This knowledge has led to the present study in which it is proved that 2.45-GHz microwaves are able to reduce ELISA incubation times. This time reduction of 50% or more concerns each of the four incubation steps of the specific indirect ELISA which is used to determine the amount of antibodies in NF90-hybridoma cell culture supernatants. These incubation steps involve the coating of the microtiter plate, the incubation with BSA against non-specific binding, and the incubations with the first and second antibody. Combinations of more microwave-influenced steps in one ELISA save a considerable amount of time. When in addition air is blown through the incubation fluid during microwaving, the extinction values are almost the same as those of conventionally performed ELISAs. Thus, a good match of power setting and irradiation time results in a total incubation time reduction from 5.5 hours to 2 hours in this particular ELISA. Carefully monitoring the temperature of the incubation fluids during irradiation with the use of a fiberoptic thermometer appeared to be of crucial importance in the development of an optimal procedure.
van Dorp
R
R
Leiden Cytology and Pathology Laboratory, The Netherlands.
Kok
P G
PG
Marani
E
E
Boon
M E
ME
Kok
L P
LP
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008872:Microwaves