Review Design of carbohydrate multiarrays V.I. Dyukova b , N.V. Shilova a , O.E. Galanina a , A.Yu. Rubina b , N.V. Bovin a, ⁎ a Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 ul. Miklukho-Maklaya, 117997, Moscow, Russian b Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 ul. Vavilova, 119991 Moscow, Russia Received 30 September 2005; received in revised form 8 December 2005; accepted 8 December 2005 Available online 5 January 2006 Abstract Recently, microarray technology has increasingly been widely applied in glycobiology. This technology has rather evident potential advantages: unlimited number of carbohydrate ligands coated onto one small sized chip, enormously low consumption of both carbohydrate ligands and carbohydrate-binding proteins to be tested, etc. Literature data demonstrate that three approaches are used for glycoarray design. The first one is based on the physical adsorption of glycomolecules on a surface (as in a common ELISA), the second one—on covalent immobilization, and the third one—on a streptavidin–biotin system. In all of the described methods, carbohydrate ligands were placed on chips as a 2D monolayer and high sensitivity was achieved due to fluorescent detection. Notably, a tendency of stepping from model chips toward real multiarrays, where the number of carbohydrate ligands can be up to two hundred, has been observed the last 2 years, this already producing a number of interesting findings when studying carbohydrate-binding proteins. In 2005 new construction, 3D glycochip was described, where 150 μm diameter polyacrylamide gel elements serve as microreactors instead of 2D dots. As a result of the 3D placement of a ligand, two orders of magnitude increase of its density is possible, this providing principal signal improvement during fluorescent detection and increasing method sensitivity. At the same time, carbohydrate consumption is low, i.e., ∼1 pmol per gel element. Copolymerization chemistry enables the immobilization of several glycomolecule classes to the gel, in particular, aminospacered oligosaccharides, polyacrylamide conjugates, and even 2-aminopyridine derivatives of oligosaccharides, which are widely used in the structural analysis of glycoprotein N-chains. © 2005 Elsevier B.V. All rights reserved. Keywords: Glycochip; Carbohydrate array; Oligosaccharide 1. 2D arrays Microarray technology allowing for the simultaneous multiparameter testing of a large number of samples at a minimal consumption of the used reagents has been increas- ingly applied widely recently for the study of biological systems, particularly in glycobiology. Several reviews have already been dedicated to glycoarrays and glycobiological problems that could be solved with their use [1–5]. Multiarray technology has rather evident advantages: unlimited (thousands) carbohydrate ligands, which can be placed on a single chip and a principal consumption decrease of carbohy- drate ligands and analytes. The latter fact can be illustrated by the work [6], where 96-well polystyrene plates were compared to a chip coated with the same polystyrene demonstrating that polysaccharide consumption at the assigned antibody concen- tration was 50 pg and 80 fg, respectively. Another advantage of multiarrays with a maximally wide glycan set should be noted: screening expands our tunnel vision and eliminates prejudice during the selection of potential ligands for particular carbohydrate-binding protein, thus providing a chance of serendipity in the search of new ligands. Such a set also removes the limitations of choice due to economic reasons: few researchers can afford purchasing one hundred or even several dozens commercial glycoconjugates for carrying out a single experiment, whereas several hundred glycans will be presented on a single chip for a reasonable price. As the key technology components, arrayers, scanners, chip platforms (microscope glass slides are usually used) and the Biochimica et Biophysica Acta 1760 (2006) 603 – 609 http://www.elsevier.com/locate/bba Abbreviations: MAb, momoclonal antibodies; A tri , GalNAcα1-3(Fucα1-2) Gal; B tri , Galα1-3(Fucα1-2)Gal; PAA, polyacrylamide; CBP, carbohydrate- binding protein; 2AP, 2-aminopyridine; Sug, saccharide residue; ELISA, enzyme-linked immunosorbent assay; All, allyl,CH 2 =CH-CH 2 -; OS, oligosaccharide ⁎ Corresponding author. Tel.: +7 495 330 71 38; fax: +7 495 330 55 92. E-mail address: bovin@carbohydrate.ru (N.V. Bovin). 0304-4165/$ - see front matter © 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.bbagen.2005.12.005