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Analytical and Bioanalytical Chemistry

, Volume 377, Issue 1, pp 220–224 | Cite as

An ELISA-based approach to optimize elution conditions for obtaining hapten-specific antibodies

  • K. V. Singh
  • Jasdeep Kaur
  • Manoj Raje
  • Grish C. Varshney
  • C. Raman SuriEmail author
Short Communication

Abstract

The correct choice of the elution conditions to break an affinity interaction is important for the successful purification of biomolecules. The optimal elution buffer liberates the bound substance in a minimum volume and maintains the activity of the purified material. The present study demonstrates an enzyme-linked immunosorbent assay (ELISA)-based approach for selection of specific elution conditions for eluting antibodies against a small molecule (atrazine) from pooled sera. Six different elution conditions were tried for the removal of antibodies from the complex. Large-scale purification of anti-atrazine antibodies from the sera was done with a hapten-specific column using an amino-terminal crosslinked agarose gel. Efficacy in terms of total amount of recovery and binding affinities of eluted antibodies from the column were further investigated by ELISA. Results indicate that the ELISA-based elution approach is ideal for the selection of suitable elution buffer that can subsequently be utilized for affinity purification applications.

Keywords

Affi-gel 102 Affinity purification Antibody ELISA Hapten 

Notes

Acknowledgements

The authors are thankful to Dr Chaaya Iyenger for the critical reviewing of the manuscript. We also greatly acknowledge the financial support of the SDC (Switzerland) and DBT (India) under their ISCB research grant.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • K. V. Singh
    • 1
  • Jasdeep Kaur
    • 1
  • Manoj Raje
    • 1
  • Grish C. Varshney
    • 1
  • C. Raman Suri
    • 1
    Email author
  1. 1.Sector 39-AInstitute of Microbial TechnologyChandigarhIndia

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