title: Emergent properties of nanosensor arrays: applications for monitoring IgG affinity distributions, weakly affined hypermannosylation, and colony selection for biomanufacturing.
abstract: It is widely recognized that an array of addressable sensors can be multiplexed for the label-free detection of a library of analytes. However, such arrays have useful properties that emerge from the ensemble, even when monofunctionalized. As examples, we show that an array of nanosensors can estimate the mean and variance of the observed dissociation constant (KD), using three different examples of binding IgG with Protein A as the recognition site, including polyclonal human IgG (KD μ = 19 μM, σ(2) = 1000 mM(2)), murine IgG (KD μ = 4.3 nM, σ(2) = 3 μM(2)), and human IgG from CHO cells (KD μ = 2.5 nM, σ(2) = 0.01 μM(2)). Second, we show that an array of nanosensors can uniquely monitor weakly affined analyte interactions via the increased number of observed interactions. One application involves monitoring the metabolically induced hypermannosylation of human IgG from CHO using PSA-lectin conjugated sensor arrays where temporal glycosylation patterns are measured and compared. Finally, the array of sensors can also spatially map the local production of an analyte from cellular biosynthesis. As an example, we rank productivity of IgG-producing HEK colonies cultured directly on the array of nanosensors itself.
authors.affiliation: Department of Chemical Engineering, Massachusetts Institute of Technology , Cambridge, Massachusetts 02139, United States.
authors.forename: Nigel F
authors.initials: NF
authors.lastname: Reuel
authors.forename: Brittany
authors.initials: B
authors.lastname: Grassbaugh
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kruss
authors.forename: J Zachary
authors.initials: JZ
authors.lastname: Mundy
authors.forename: Cary
authors.initials: C
authors.lastname: Opel
authors.forename: Adebola O
authors.initials: AO
authors.lastname: Ogunniyi
authors.forename: Kamal
authors.initials: K
authors.lastname: Egodage
authors.forename: Ramon
authors.initials: R
authors.lastname: Wahl
authors.forename: Bernhard
authors.initials: B
authors.lastname: Helk
authors.forename: Jingqing
authors.initials: J
authors.lastname: Zhang
authors.forename: Z Ilke
authors.initials: ZI
authors.lastname: Kalcioglu
authors.forename: Kevin
authors.initials: K
authors.lastname: Tvrdy
authors.forename: Darin O
authors.initials: DO
authors.lastname: Bellisario
authors.forename: Bin
authors.initials: B
authors.lastname: Mu
authors.forename: Steven S
authors.initials: SS
authors.lastname: Blake
authors.forename: Krystyn J
authors.initials: KJ
authors.lastname: Van Vliet
authors.forename: J Christopher
authors.initials: JC
authors.lastname: Love
authors.forename: Karl Dane
authors.initials: KD
authors.lastname: Wittrup
authors.forename: Michael S
authors.initials: MS
authors.lastname: Strano
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques / Q000295:instrumentation*; D001681:Biological Assay / Q000295:instrumentation*; D015374:Biosensing Techniques / Q000295:instrumentation*; D016466:CHO Cells; D003114:Colony-Forming Units Assay / Q000295:instrumentation; D003412:Cricetulus; D004867:Equipment Design; D019544:Equipment Failure Analysis; D057809:HEK293 Cells; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D007074:Immunoglobulin G / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D008358:Mannose / Q000737:chemistry / Q000276:immunology; D051379:Mice; D037742:Nanotubes, Carbon / Q000737:chemistry* / Q000648:ultrastructure; D011485:Protein Binding; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000276:immunology
chemical_list: D007074:Immunoglobulin G; D037742:Nanotubes, Carbon; D013205:Staphylococcal Protein A; D008358:Mannose