Immunoaffinity purification of indole-3-acetamide using monoclonal antibodies
Research output: Contribution to journal › Journal article › Research › peer-review
A single-step immunoaffinity purification procedure using monoclonal antibodies was developed to isolate indole-3-acetamide from plant extracts. Antibodies from a selected clone, raised against IAA-Cl'-BSA, with pronounced ability to recognize indole-3-acetamide (IAM) were used to prepare an immunoaffinity absorbent. Antibodies purified by thiophilic interaction chromatography were immobilized on divinylsulfoneactivated agarose. This column shows a very high selectivity towards IAM compared to IAA. This single step of immunoaffinity purification gave plant extracts of sufficient purity for direct quantification by on-line spectrofluorimetry after an analytical ionsuppression-HPLC run. Successive approximation by a second analytical ion-pairing-HPLC run confirmed the validity of this analytical technique.
| Original language | English |
|---|---|
| Journal | Plant and Cell Physiology |
| Volume | 28 |
| Issue number | 5 |
| Pages (from-to) | 937-945 |
| Number of pages | 9 |
| ISSN | 0032-0781 |
| DOIs | |
| Publication status | Published - Jul 1987 |
Bibliographical note
Funding Information:
Britta Nielsen is acknowledged for her skillful technical assistance. This work wassupported by the Danish Sugar Factories (Peter Ulvskov) and by the Danish Agricultural Research Council toJan Marcussen, vide J.nrs.: SJVF 13-3224 and 13-3406. Els Prinsen is recipient of an "Instituut voor Wetenschappelijk Onderzoek in Nijverheid en Landbouw" (IWONL) grant. Patrick Rudelsheim and Henri Van Onckelen are research assistant and senior research associate respectively of "Nationaal Fonds voor Wetenschappelijk Onderzoek" (NFWO).
- HPLC-on-line-fluorimetry, Immunoaffinity chromatography, Indole-3-acetamine, T-DNA, Thiophilic interaction chromatography, Tritiated indole-3-acetamide
Research areas
ID: 308328443