Immunoaffinity purification of indole-3-acetamide using monoclonal antibodies
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Immunoaffinity purification of indole-3-acetamide using monoclonal antibodies. / Ulvskov, Peter; Marcussen, Jan; Rajagopal, Ranganatha; Prinsen, Els; Rudelsheim, Patrick; Onckelen, Henri Van.
In: Plant and Cell Physiology, Vol. 28, No. 5, 07.1987, p. 937-945.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Immunoaffinity purification of indole-3-acetamide using monoclonal antibodies
AU - Ulvskov, Peter
AU - Marcussen, Jan
AU - Rajagopal, Ranganatha
AU - Prinsen, Els
AU - Rudelsheim, Patrick
AU - Onckelen, Henri Van
N1 - 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).
PY - 1987/7
Y1 - 1987/7
N2 - 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.
AB - 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.
KW - HPLC-on-line-fluorimetry
KW - Immunoaffinity chromatography
KW - Indole-3-acetamine
KW - T-DNA
KW - Thiophilic interaction chromatography
KW - Tritiated indole-3-acetamide
UR - http://www.scopus.com/inward/record.url?scp=70349864415&partnerID=8YFLogxK
U2 - 10.1093/oxfordjournals.pcp.a077376
DO - 10.1093/oxfordjournals.pcp.a077376
M3 - Journal article
AN - SCOPUS:70349864415
VL - 28
SP - 937
EP - 945
JO - Plant and Cell Physiology
JF - Plant and Cell Physiology
SN - 0032-0781
IS - 5
ER -
ID: 308328443