A comprehensive toolkit of plant cell wall glycan-directed monoclonal antibodies.
A collection of 130 new plant cell wall glycan-directed monoclonal antibodies (mAbs) was generated with the aim of facilitating in-depth analysis of cell wall glycans. An enzyme-linked immunosorbent assay-based screen against a diverse panel of 54 plant polysaccharides was used to characterize the binding patterns of these new mAbs, together with 50 other previously generated mAbs, against plant cell wall glycans. Hierarchical clustering analysis was used to group these mAbs based on the polysaccharide recognition patterns observed. The mAb groupings in the resulting cladogram were further verified by immunolocalization studies in Arabidopsis (Arabidopsis thaliana) stems. The mAbs could be resolved into 19 clades of antibodies that recognize distinct epitopes present on all major classes of plant cell wall glycans, including arabinogalactans (both protein- and polysaccharide-linked), pectins (homogalacturonan, rhamnogalacturonan I), xyloglucans, xylans, mannans, and glucans. In most cases, multiple subclades of antibodies were observed to bind to each glycan class, suggesting that the mAbs in these subgroups recognize distinct epitopes present on the cell wall glycans. The epitopes recognized by many of the mAbs in the toolkit, particularly those recognizing arabinose- and/or galactose-containing structures, are present on more than one glycan class, consistent with the known structural diversity and complexity of plant cell wall glycans. Thus, these cell wall glycan-directed mAbs should be viewed and utilized as epitope-specific, rather than polymer-specific, probes. The current world-wide toolkit of approximately 180 glycan-directed antibodies from various laboratories provides a large and diverse set of probes for studies of plant cell wall structure, function, dynamics, and biosynthesis.
Pattathil
Sivakumar
S
Complex Carbohydrate Research Center , University of Georgia, Athens, Georgia 30602, USA.
Avci
Utku
U
Baldwin
David
D
Swennes
Alton G
AG
McGill
Janelle A
JA
Popper
Zoë
Z
Bootten
Tracey
T
Albert
Anathea
A
Davis
Ruth H
RH
Chennareddy
Chakravarthy
C
Dong
Ruihua
R
O'Shea
Beth
B
Rossi
Ray
R
Leoff
Christine
C
Freshour
Glenn
G
Narra
Rajesh
R
O'Neil
Malcolm
M
York
William S
WS
Hahn
Michael G
MG
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002473:Cell Wall / Q000737:chemistry*; D016000:Cluster Analysis; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D010944:Plants / Q000737:chemistry*; D011134:Polysaccharides / Q000032:analysis*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011134:Polysaccharides