title: Printing low density protein arrays in microplates.
abstract: Here, we provide methods for the creation of protein microarrays in microplates. The microplate consists of 96 wells with each well capable of holding a protein microarray at a spot density of up to 400 (20 x 20) individual elements. Arrays of capture monoclonal antibodies, corresponding to specific interleukins, were printed onto the bottom of the wells which had been surface activated for covalent attachment. A Biomek 2000 laboratory automation workstation (Beckman Coulter, Inc., Fullerton, CA) equipped with a high-density replicating tool was used for printing low density 3 x 3 to 5 x 5 arrays. For higher density arrays, a microarrayer system (Cartesian PS7200, Genomic Solutions, Inc., Ann Arbor, MI) was employed. Multiple antigens were simultaneously analyzed without detectable cross-reactivity associated with capture antibody or secondary antibody interactions. Detection of interleukin antigens, spiked into cell culture media containing 10% fetal calf serum, was specific and sensitive.
authors.affiliation: Beckman Coulter, Inc., Fullerton, CA, USA.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Matson
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Milton
authors.forename: Michael C
authors.initials: MC
authors.lastname: Cress
authors.forename: Tom S
authors.initials: TS
authors.lastname: Chan
authors.forename: Jang B
authors.initials: JB
authors.lastname: Rampal
mesh_terms: D000818:Animals; D000906:Antibodies; D016207:Cytokines / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods*; D012984:Software
chemical_list: D000906:Antibodies; D016207:Cytokines