Microbead analysis of cell binding to immobilized lectin: an alternative to microarrays in the development of carbohydrate drugs and diagnostic tests.
Microarray technology is currently used in the development of carbohydrate drugs and diagnostic tests. Here we model an inexpensive alternative to microarrays using derivatized microbeads. In this model we examine the binding of mannose-rich yeast to microbeads derivatized with concanavalin A (Con A), a mannose-binding lectin, in the presence of 30 different sugars and 9 different pH conditions. We developed a listing of effective saccharide inhibitors of immobilized Con A based on 3901 replicates. We suggest that this is the most extensive saccharide inhibitor list ever developed for this lectin and it may be useful to use this listing to replace the less extensive lists that have been in the literature for decades. Information is also provided on pH effects on immobilized Con A binding based on 918 trials. Two assays to study binding, one which qualitatively scores more or less binding than control in thousands of replicate samples, and another that quantitatively evaluates binding by counting the number of cells bound to each bead, are also modeled here. We know of no previous studies that provide such extensive information on saccharide inhibition and pH effects on the binding of immobilized Con A. We suggest that this microbead approach, using beads derivatized with lectins or sugars, and the two simple assays presented here, can in some cases substitute for more expensive microarray technology in the development of carbohydrate drugs and diagnostic tests. If, for example, our model Saccharomyces cerevisiae was a pathogen, these studies show that it binds via cell surface mannose residues and drugs to prevent binding could be developed using the inhibitors of binding identified here. The beads could be also used in the development of diagnostic tests that identify the presence of the organism in blood samples, etc. in much the same way as microarray technology is being used today.
Zem
Gregory C
GC
Department of Biology and Center for Cancer and Developmental Biology, California State University Northridge, 18111 Nordhoff St. Northridge, CA 91330-8303, USA.
Badali
Oliver
O
Gaytan
Maria
M
Hekmatjou
Hesam
H
Alvarez
Maribel
M
Nnoli
Jennifer
J
Katus
Elena
E
Oppenheimer
Steven B
SB
D002241:Carbohydrates / Q000494:pharmacology* / Q000627:therapeutic use; D019937:Diagnostic Techniques and Procedures; D015195:Drug Design; D005057:Eukaryotic Cells / Q000378:metabolism*; D037102:Lectins / Q000737:chemistry*; D046228:Microarray Analysis; D008863:Microspheres; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013499:Surface Properties
D002241:Carbohydrates; D037102:Lectins