Antibody mediated fluorescence enhancement of nucleoside analogue 1,3-diaza-2-oxophenoxazine (tC°)

• Generation of antibody against the tricyclic fluorescent nucleoside analogue 1,3-diaza-2-oxophenoxazine (tC°).
• Antibody mediated fluorescence enhancement and hypsochromic shift of fluorophore tC°.
• Application of tC° binding antibody in homogeneous immunoassay for the sequence specific determination of DNA.

We report on the generation and analytical application of the monoclonal antibody G93-ED2 raised against the tricyclic fluorescent nucleoside analogue 1,3-diaza-2-oxophenoxazine (tC°). G93-ED2 is specifically binding this deoxycytidine analogue and was found to raise its fluorescence intensity by a factor of 5. This unique feature makes it a valuable tool in fluorescence dependent immunoassays. G93-ED2 was successfully applied in a homogeneous fluorescence quenching immunoassay (DNA-Q) for the sequence specific determination of DNA.

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