An Antibody Fab Selected from a Recombinant Phage Display Library Detects Deesterified Pectic Polysaccharide Rhamnogalacturonan II in Plant Cells

Rhamnogalacturonan II (RG-II) is a structurally complex, low molecular weight pectic polysaccharide that is released from primary cell walls of higher plants by treatment with endopolygalacturonase and is chromatographically purified after alkaline deesterification. A recombinant monovalent antibody...

Full description

Saved in:
Bibliographic Details
Published in:The Plant cell 1996-04, Vol.8 (4), p.673-685
Main Authors: Myron N. V. Williams, Freshour, Glenn, Darvill, Alan G., Albersheim, Peter, Hahn, Michael G.
Format: Article
Language:English
Subjects:
Animals
Antibodies
Bacteriophages
Base Sequence
Cell membranes
Cell walls
Coliphages
DNA Primers - chemistry
Epitopes
Fluorescent Antibody Technique, Indirect
Gene Library
Genes, Immunoglobulin
Immunoglobulin Fab Fragments - chemistry
Immunohistochemistry
Libraries
Mice
Mice, Inbred BALB C
Molecular Sequence Data
Monoclonal antibodies
Pectins - chemistry
Pectins - immunology
Plant cells
Plants
Polysaccharides
Rhamnose
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Rhamnogalacturonan II (RG-II) is a structurally complex, low molecular weight pectic polysaccharide that is released from primary cell walls of higher plants by treatment with endopolygalacturonase and is chromatographically purified after alkaline deesterification. A recombinant monovalent antibody fragment (Fab) that specifically recognizes RG-II has been obtained by selection from a phage display library of mouse immunoglobulin genes. By itself, RG-II is not immunogenic. Therefore, mice were immunized with a neoglycoprotein prepared by covalent attachment of RG-II to modified BSA. A cDNA library of the mouse IgG1/κ antibody repertoire was constructed in the phage display vector pComb3. Selection of antigen-binding phage particles resulted in the isolation of an antibody Fab, CCRC-R1, that binds alkali-treated RG-II with high specificity. CCRC-R1 binds an epitope found primarily at sites proximal to the plasma membrane of suspension-cultured sycamore maple cells. In cells deesterified by alkali, CCRC-R1 labels the entire wall, suggesting that the RG-II epitope recognized by CCRC-R1 is masked by esterification in most of the wall and that such RG-II esterification is absent near the plasma membrane.
ISSN:1040-4651
1532-298X
DOI:10.1105/tpc.8.4.673