The purpose of this study was to produce a highly specific monoclonal antibody against free indole-3-acetic acid (IAA) and to develop a competitive enzyme linked immunosorbent assay (CELIA). Hybridomas were produced by fusion of NS 1 myeloma cells with spleen cells from BALB/cj mice immunized with IAA conjugated to rat serum albumin via the indole nitrogen. A highly specific cell line secreting IgG 1 chain specific kappa light chain for IAA was obtained. This cell line has little or no cross reactivity with 2,4-dichlorophenoxyacetic acid (2,4-D) and structurally related indole compounds (including amino-acid conjugates). The standard competitive ELISA assay provides sensitivity in the pmol range. Crown gall tumor derived from mung bean (Vigna radiata cv. Berken) stem tissue extracts were passed through SepPak β-Bondapak C-18 cartridges, Sephadex G-10, carboxymethyl-cellulose (CMC), polyvinylpolypyr-rolidone (PVPP) and DEAE-cellulose columns individually and in combination in order to evaluate what degree of purification was required in order to avoid interference with the immunological assay. It was found that SepPak and PVPP in combination provided an adequate clean-up for use with plant samples.
All Science Journal Classification (ASJC) codes
- Agronomy and Crop Science
- Plant Science