BIST 3/3

We demonstrate the applications of BIST technology in multiplex SNP analysis, infectious disease testing and in
multiplex ELISA, all incorporating this novel automation system. The BIST System has performed with 100%
concordant results for 4 SNPs commonly associated with obesity-related analysis. The BIST assay that was
developed is contained with in a single tip of 18 immobilized beads representing nine different tagged oligo DNA and
quenchers. These tagged oligos have been developed for isothermal hybridization, maintaining unique sequences.
The possibility of a multiplex ELISA exists incorporating beads immobilized with anti-AFP antibody and anti-Albumin
antibody.

1. Coated antigen A and B on the beads and made 2 kinds of BIST.
2. Reacted the positive plasma diluted to 1/614400-1/2400 in antigen A
BIST, to 1/12800-1/50 in antigen B BIST using Magtration system .
3. Reacted with anti Human IgG, HRP as a second antibody.
4. Luminescence intensity measured with BISTnner.

1. Beads were coated with anti AFP antibody and anti albumin antibody.
2. Two sets of three beads are placed to create BIST capillary (Figure drawing)
3. Preparation of reaction mix containing antigen and antibody, along with POD
and HRP, are prepared, dispensed and hybridized on Magtration system .
4. Luminescence intensity is measured with the BISTnner.

We were able to confirm linearity in concentrations as low as 1/76800 for antigen A ,
and in less than a 1/200 concentration for antigen B. Using the BIST system, we were able
to detect equal or lower concentrations than that with ELISA plate detection

Both AFP and Albumin can be detected with BIST. Antigen
concentrations of 0.5-250ng/ml can be detect ed in antigen
mixture. This result shows the possibility of a multiplex ELISA