Emergent properties of nanosensor arrays: applications for monitoring IgG affinity distributions, weakly affined hypermannosylation, and colony selection for biomanufacturing

ACS Nano. 2013 Sep 24;7(9):7472-82. doi: 10.1021/nn403215e. Epub 2013 Aug 15.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Batch Cell Culture Techniques / instrumentation*
  • Biological Assay / instrumentation*
  • Biosensing Techniques / instrumentation*
  • CHO Cells
  • Colony-Forming Units Assay / instrumentation
  • Cricetulus
  • Equipment Design
  • Equipment Failure Analysis
  • HEK293 Cells
  • Humans
  • Immunoassay / instrumentation*
  • Immunoglobulin G / analysis*
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / immunology
  • Mannose / chemistry
  • Mannose / immunology
  • Mice
  • Nanotubes, Carbon / chemistry*
  • Nanotubes, Carbon / ultrastructure
  • Protein Binding
  • Staphylococcal Protein A / chemistry
  • Staphylococcal Protein A / immunology

Substances

  • Immunoglobulin G
  • Nanotubes, Carbon
  • Staphylococcal Protein A
  • Mannose