10. A Novel Multiplex SPR Array for Rapid Screening and Affinity Determination of Monoclonal Antibodies: The ProteOn XPR36 Label Free System: Kinetic Screening of Monoclonal Antibodies

  1. Matthew Cooper2 and
  2. Lorenz M. Mayr3
  1. Vered Bronner,
  2. Oded Nahshol and
  3. Tsafrir Bravman

Published Online: 24 FEB 2011

DOI: 10.1002/9780470979129.ch10

Label-Free Technologies for Drug Discovery

Label-Free Technologies for Drug Discovery

How to Cite

Bronner, V., Nahshol, O. and Bravman, T. (2011) A Novel Multiplex SPR Array for Rapid Screening and Affinity Determination of Monoclonal Antibodies: The ProteOn XPR36 Label Free System: Kinetic Screening of Monoclonal Antibodies, in Label-Free Technologies for Drug Discovery (eds M. Cooper and L. M. Mayr), John Wiley & Sons, Ltd, Chichester, UK. doi: 10.1002/9780470979129.ch10

Editor Information

  1. 2

    Institute for Molecular Bioscience, University of Queensland, Australia

  2. 3

    Biology Unit, Protease Platform, Novartis Pharma AG, Basel, Switzerland

Author Information

  1. Bio-Rad Laboratories, Inc., Gutwirth Park, Technion, Haifa 32000, Israel

Publication History

  1. Published Online: 24 FEB 2011
  2. Published Print: 25 FEB 2011

ISBN Information

Print ISBN: 9780470746837

Online ISBN: 9780470979129

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Keywords:

  • Novel Multiplex SPR Array for rapid screening - affinity determination of monoclonal antibodies, the ProteOn XPR36 Label Free System;
  • Surface Plasmon Resonance (SPR) optical biosensing - either radiochemical or fluorescent labels to provide real-time data on affinity;
  • one-shot parallel approach - complete kinetic profile of biomolecular interaction;
  • optimized assay configuration - kinetic binding and affinity constants, for hundreds of monoclonal supernatants raised against two antigens;
  • workflow for kinetic analysis of antibodies supernatants;
  • optimal capture agent, meeting criteria - ability to bind to sensor chip in high density;
  • capture agents, anti-mouse IgG whole molecule - protein A/G; anti-mouse IgG Fc specific, and anti-mouse Ig;
  • crucial parameter - preservation of captured antibody activity for antigen binding;
  • interspot references - in horizontal and vertical channel flow path of sensor chip;
  • kinetic analysis of 243 human hemoglobin supernatants

Summary

This chapter contains sections titled:

  • Introduction

  • Optimized Assay Configuration

  • Selection of the Optimal Capture Agent

  • Kinetic Analysis of 192 Human Anti-IL-12 Supernatants

  • Kinetic Analysis of 243 Human Hemoglobin Supernatants

  • Conclusions

  • References