earticle

영어

The expression of metallothionein in O.javanicus (OjaMT) increases as a result of heavy metal exposure. Measuring the level of OjaMT expression in O.javanicus can be a good monitoring method for heavy metal contamination of environment and their toxicity. Previously, western blotting and ELISA methods were used to monitor the level of OjaMT expression and these methods requires long detection time and gives high level of background. Also, it was rather difficult to achieve quantitative measurement of OjaMT. Here, we set-up an assay to quantitatively detect OjaMT using surface plasmon resonance (SPR) system. We have used OjaMT specific antibody for the SPR based label free detection of OjaMT. Also, we have constructed OjaMT specific scFv using the total RNA of antibody generating hybridoma cells using jumping PCR. The generated scFv’s exhibited specific binding activity to OjaMT in solution. ScFv’s were immobilized to gold substrate using a 3-Cys tag introduced during the cloning vector construction. The performance of two different probes were compared and the amount of OjaMT was measured quantitatively.