title: Generation of miniaturized planar ecombinant antibody arrays using a microcantilever-based printer.
abstract: Miniaturized (Ø 10 μm), multiplexed (>5-plex), and high-density (>100 000 spots cm(-2)) antibody arrays will play a key role in generating protein expression profiles in health and disease. However, producing such antibody arrays is challenging, and it is the type and range of available spotters which set the stage. This pilot study explored the use of a novel microspotting tool, Bioplume(TM)-consisting of an array of micromachined silicon cantilevers with integrated microfluidic channels-to produce miniaturized, multiplexed, and high-density planar recombinant antibody arrays for protein expression profiling which targets crude, directly labelled serum. The results demonstrated that 16-plex recombinant antibody arrays could be produced-based on miniaturized spot features (78.5 um(2), Ø 10 μm) at a 7-125-times increased spot density (250 000 spots cm(-2)), interfaced with a fluorescent-based read-out. This prototype platform was found to display adequate reproducibility (spot-to-spot) and an assay sensitivity in the pM range. The feasibility of the array platform for serum protein profiling was outlined.
authors: Department of Immunotechnology, Lund University, Medicon Village, SE-22381 Lund, Sweden. CREATE Health, Lund University, Medicon Village, SE-22381 Lund, Sweden.
authors: Linn
authors: L
authors: Petersson
authors: Nathalie
authors: N
authors: Berthet Duroure
authors: Angèle
authors: A
authors: Auger
authors: Linda
authors: L
authors: Dexlin-Mellby
authors: Carl Ak
authors: CA
authors: Borrebaeck
authors: Ali
authors: A
authors: Ait Ikhlef
authors: Christer
authors: C
authors: Wingren
mesh_terms: D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D001798:Blood Proteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods*; D044085:Microfluidics / Q000295:instrumentation; D008904:Miniaturization; D010865:Pilot Projects; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology
chemical_list: D000906:Antibodies; D001798:Blood Proteins; D011994:Recombinant Proteins