title: Multiparameter Particle Display (MPPD): A Quantitative Screening Method for the Discovery of Highly Specific Aptamers.
abstract: Aptamers are a promising class of affinity reagents because they are chemically synthesized, thus making them highly reproducible and distributable as sequence information rather than a physical entity. Although many high-quality aptamers have been previously reported, it is difficult to routinely generate aptamers that possess both high affinity and specificity. One of the reasons is that conventional aptamer selection can only be performed either for affinity (positive selection) or for specificity (negative selection), but not both simultaneously. In this work, we harness the capacity of fluorescence activated cell sorting (FACS) for multicolor sorting to simultaneously screen for affinity and specificity at a throughput of 107aptamers per hour. As a proof of principle, we generated DNA aptamers that exhibit picomolar to low nanomolar affinity in human serum for three diverse proteins, and show that these aptamers are capable of outperforming high-quality monoclonal antibodies in a standard ELISA detection assay.
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: Jinpeng
authors.initials: J
authors.lastname: Wang
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: Jingwen
authors.initials: J
authors.lastname: Yu
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: Qin
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: John
authors.initials: J
authors.lastname: McDermott
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Scott
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Vukovich
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: Remy
authors.initials: R
authors.lastname: Lagrois
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Gong
authors.affiliation: Department of Genetics, Stanford University, Stanford, CA, 94305, USA.
authors.forename: William
authors.initials: W
authors.lastname: Greenleaf
authors.affiliation: Department of Electrical Engineering and Radiology, Stanford University, Stanford, CA, 94305, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Eisenstein
authors.affiliation: Aptitude Medical Systems Inc., Santa Barbara, CA, 93105, USA.
authors.forename: B Scott
authors.initials: BS
authors.lastname: Ferguson
authors.affiliation: Department of Electrical Engineering and Radiology, Stanford University, Stanford, CA, 94305, USA.
authors.forename: H Tom
authors.identifier: 0000-0001-9443-857X
authors.initials: HT
authors.lastname: Soh
mesh_terms: D052157:Aptamers, Nucleotide / Q000097:blood* / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D010316:Particle Size
chemical_list: D052157:Aptamers, Nucleotide
keywords: affinity reagents; aptamers; flow cytometry; high-throughput screening; molecular recognition