creators_name: Singh, K V
creators_name: Kaur, Jasdeep
creators_name: Raje, Manoj
creators_name: Varshney, Grish C
creators_name: Suri, C Raman
type: article
datestamp: 2012-01-06 14:58:10
lastmod: 2012-01-06 14:58:10
metadata_visibility: show
title: An ELISA-based approach to optimize elution conditions for obtaining hapten-specific antibodies.
ispublished: pub
subjects: QD
full_text_status: restricted
keywords: Affi-gel 102 · Affinity purification ·
Antibody · ELISA · Hapten
note: Copyright of this article belongs to Springer Science.
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.
date: 2003-09
date_type: published
publication: Analytical and bioanalytical chemistry
volume: 377
number: 1
publisher: Springer Science
pagerange: 220-4
refereed: TRUE
issn: 1618-2642
official_url: http://www.springerlink.com/content/ubrl548pjw5xrrte/fulltext.pdf
citation:   Singh, K V and Kaur, Jasdeep and Raje, Manoj and Varshney, Grish C and Suri, C Raman  (2003) An ELISA-based approach to optimize elution conditions for obtaining hapten-specific antibodies.  Analytical and bioanalytical chemistry, 377 (1).  pp. 220-4.  ISSN 1618-2642     
document_url: http://crdd.osdd.net/open/239/1/suri2003.pdf