A radioimmunoassay for lignin in plant cell walls
Lignin detection and determination in herbaceous tissue requires selective, specific assays which are not currently available. A radioimmunoassay (RIA) was developed to study lignin metabolism in these tissues. A {beta}-aryl ether lignin model compound was synthesized, linked to keyhole limpet hemocyanin using a water-soluble carbodiimide, and injected into rabbits. The highest titer of the antiserum obtained was 34 {eta}g/mL of model derivatized BSA. An in vitro system was developed to characterize the RIA. The model compound was linked to amino activated polyacrylamide beads to mimic lignin in the cell walls. {sup 125}I Radiolabelled protein A was used to detect IgG antibody binding. The RIA was shown in the in vitro system to exhibit saturable binding. The amount of antibody bound decreased when the serum was diluted. Immunoelectrophoresis and competitive binding experiments confirmed that both aromatic rings of the lignin model compound had been antigenic. Chlorogenic acid, a phenolic known to be present in plant cells, did not compete for antibody binding. The RIA was used to measure lignin in milled plant samples and barley seedlings. Antiserum binding to wheat cell walls and stressed barley segments was higher than preimmune serum binding. Antibody binding to stressed barley tissue decreased following NaClO{sub 2} delignification. The RIA was found to be less sensitive than expected, so several avenues for improving the method are discussed.
- Research Organization:
- Miami Univ., Coral Gables, FL (USA)
- OSTI ID:
- 5730258
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
09 BIOMASS FUELS
LIGNIN
RADIOIMMUNOASSAY
BARLEY
BIOCHEMICAL REACTION KINETICS
CELL WALL
CPB
ELECTROPHORESIS
IODINE 125
METABOLISM
WHEAT
BETA DECAY RADIOISOTOPES
BIOASSAY
CARBOHYDRATES
CELL CONSTITUENTS
CEREALS
DAYS LIVING RADIOISOTOPES
DIAGNOSTIC TECHNIQUES
ELECTRON CAPTURE RADIOISOTOPES
GRAMINEAE
IMMUNOASSAY
IMMUNOLOGY
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LILIOPSIDA
MAGNOLIOPHYTA
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PLANTS
POLYSACCHARIDES
RADIOASSAY
RADIOIMMUNODETECTION
RADIOIMMUNOLOGY
RADIOISOTOPES
REACTION KINETICS
SACCHARIDES
TRACER TECHNIQUES
550501* - Metabolism- Tracer Techniques
090800 - Biomass Fuels- Production- (1990-)