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

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

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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.

Item Type:	Article
Additional Information:	Copyright of this article belongs to Springer Science.
Uncontrolled Keywords:	Affi-gel 102 · Affinity purification · Antibody · ELISA · Hapten
Subjects:	Q Science > QD Chemistry
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	06 Jan 2012 14:58
Last Modified:	06 Jan 2012 14:58
URI:	http://crdd.osdd.net/open/id/eprint/239

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