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Article Dans Une Revue Analytical Biochemistry Année : 2008

Block copolymer-oligonucleotide conjugates for genotyping on microarrays

Résumé

Polymer-oligonucleotide conjugates were synthesized from the amphiphilic block copolymer poly(tern -butylacrylamide-b -(N -acryl-oylmorpholine-co-N-acryloxysuccinimide)) using an original solid-phase DNA synthesis strategy. This method provided conjugates highly functionalized with oligonucleotides throughout the polymer chain. After purification, block copolymer-oligonucleotide conjugates were spotted on a multidetection microarray system developed by Apibio using a standard nanodroplet piezo inkjet spotting technique to develop the oligosorbent assay (OLISA). Two genotyping models (HLA-DQB1 and platelet glycoproteins [GPs]), which are particularly difficult to study with standard systems, were evaluated. For both models, block copolymer-oligonucleotide conjugates used as capture probes amplified the responses of in vitro diagnostic assays. The detection limit reached by using conjugates was estimated at 15 pM for a 219-bp DNA target (HLA-DQB1 model). Moreover, single nucleotide polymorphism was detected in the platelet GPs genotyping model. The use of polymer conjugates led to a significant improvement in both sensitivity and specificity of standard hybridization assays even when applied to complex biological models.

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Dates et versions

hal-00373767 , version 1 (07-04-2009)

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  • HAL Id : hal-00373767 , version 1

Citer

Bertrand de Lambert, Carole Chaix, Marie-Thérèse Charreyre, Thibault Martin, Arnaud Aigoui, et al.. Block copolymer-oligonucleotide conjugates for genotyping on microarrays. Analytical Biochemistry, 2008, 373 (2), pp.229-238. ⟨hal-00373767⟩
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