High Throughput Filtering Assays for Multiplexed Blood Group Genotyping and Human Serology – HIFI-ASSAYS

The proposed technology is to use an innovative filtration array (HiFi) together with cost-efficient colorimetric staining/detection of the positive results (Patent No. FR09356049.8 and US61/227 666) . In this concept, different molecular probes can be immobilised as medium density (100 spots) microarrays (HiFi-array) on the external face of a microtiter plate bottom modified with a porous polymeric membrane The samples are then incubated/filtrated through the plate bottom, optimising the interaction between target and probe molecules.

Some technological bottlenecks have been broken to move forward. We have changed the format of microtitter plate and modified some robotic gears to allow better handling. We now have very encouraging results.

Blood genotyping application is highly promising and we are currently working on a pilot program on the EFS premises. This wil allow validation of commercial kit at industrial level.

Revues à comité de lecture
1. Mandon C.A., Berthuy O.I., Corgier B.P., Le Goff G.C., Faure P., Marche P.N., Blum L.J. and Marquette C.A., Polymer adhesive surface as flexible generic platform for multiplexed assays biochip production. Biosensors & Bioelectronics, in press.

Communications (conférence)
1. B.P. Corgier, Adhesive Microarrays: A New Integrative Solution. Advance Microarray technology, 28th – 29th march 2012, Edinburgh (Scotland). Oral Communication.
2. Benjamin P. Corgier, Céline A. Mandon, Gaëlle C. Le Goff, Patrice Marche, Loïc J. Blum and Christophe A. Marquette (2012), Polymer adhesive surface as flexible generic platform for biochip production. Biosensors 2012, 16th - 19th May, Cancun (Mexico). Oral Communication.

The HiFi-assays project aims at developing high throughput and multiparametric analysis tools dedicated to the characterisation of biological samples. The proposed technology is to use an innovative filtration array (HiFi) together with a cost-efficient colorimetric staining/detection of positive results (Patent No. FR09356049.8 and US61/227 666). In this concept, different molecular probes can be immobilised as medium density (100 spots) microarrays (HiFi-array) on the external face of a microtiter plate bottom modified with a porous polymeric membrane. The samples are then incubated/filtrated through the plate bottom, optimising the interaction between target and probe molecules.
The plus of the technology are:
- A generic platform for multiplexed high throughput assays (DNA, protein and peptide based assays).
- A quantitative detection of interactions levels on each spots (image greyscale quantification over three decades).
- A filtering protocol increasing interactions between targets and immobilised probes and lowering the assay background.

Two applications of the innovative technology, with strong impact onto health security, will be investigated during the project:
- A DNA microarray genotyping assay for extended blood group systems characterisation of blood donation samples.
- A proteins and peptides microarray based test for the serological investigation of hepatitis infection (diagnosis) and evolution to liver cancer (prognosis).
In both case, the project will aim at developing a complete solution, from sample preparation (DNA extraction and PCR multiplex, for example) to protocol automation and treatment of data results.
Each application will be validated at medium scale (between 1’000 and 10’000 samples), during a specific on-site demonstration/validation task.

The project is an industrial project, coordinated by the company AXO Science SAS, aiming at the maturation of a product meeting the In Vitro Diagnosis (IVD) standards.
As a final deliverable and thanks to coordination by the supporting SME, a final product prototype named HiFi will be produced at a pre-industrial scale and used for project validation. Two different products named HiFi Blood 96 and HiFi Sero 96 (Trademarks already registered at the INPI), for blood genotyping and hepatitis serology, respectively, are planned to be developed following the project completion. Here, the presence in the consortium of two end-users (Etablissement Français du Sang and Institut Albert Bonniot INSERM-UJF U823) will strongly increase the project impact.