We are developing ELISAs of steroid hormones wherein antibody is immobilised on microtitre plate, the labelled steroid compete with unlabelled steroid in assay system. When same format is transferred to lateral flow assay the sensitivity goes down 100 to 500 times why?
Lateral flow immunoassays usually have a relative short incubation times especially in the area where the antibodies and the antigen can be in contact with each other. Also, if you are working with lateral flow in combination with nitrocellulose membranes you do not have that many options to modify the incubation time except increasing the length of the nitrocellulose strip.
My impression is that a plastic surface in combination with a micro-pillar flow technique gives more control of the flow and if you look at the troponin I assay developed by fiomi (www.fiomi.se/) it has a sensitivity very close to high sensitivity troponin assays. The Fiomi technology uses plastic surfaces so the technology is probably more similar to your original ELISA than the lateral flow assays that you are working with. The plastic surface also allows for more effective washing than nitrocellulose.
Lateral flow immunoassays usually have a relative short incubation times especially in the area where the antibodies and the antigen can be in contact with each other. Also, if you are working with lateral flow in combination with nitrocellulose membranes you do not have that many options to modify the incubation time except increasing the length of the nitrocellulose strip.
My impression is that a plastic surface in combination with a micro-pillar flow technique gives more control of the flow and if you look at the troponin I assay developed by fiomi (www.fiomi.se/) it has a sensitivity very close to high sensitivity troponin assays. The Fiomi technology uses plastic surfaces so the technology is probably more similar to your original ELISA than the lateral flow assays that you are working with. The plastic surface also allows for more effective washing than nitrocellulose.
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