Recombinant Prokaryotic Lectins

Lectins can be immobilized onto a wide range of solid supports, creating glycoselective bioaffinity matrices. By doing this, lectins are enabled to recognise and bind to specific glycans. This exploits the isolation and separation of glycosylated biomolecules and their glycoforms based on the glycans they display and overcomes limitations of traditional separation tools and techniques that work on the bases of size, charge status or hydrophobicity/hydrophilicity.

AMSBIO offers Recombinant prokaryotic lectins (RPL), which are superior glycoselective biaffinity molecules that enable simple, fast, and efficient detection, analysis, and isolation of intact glycosylated biomolecules for your research.

• Specific, high-affinity glycoselective binding
• Simple and scalable production
• Consistent high quality
• Superior performance
• Contain poly-histidine tag
• No disulphide bonds

Currently, most commercially available lectins are from plants and other eukaryotes. Whilst these offer many advantages as glycoanalytical tools, they still carry with them many limitations:

• Lack of selectivity - Plant derived lectins are often reported to bind to a range of different glycan structures and so lack the selectivity required for many applications.
• Inconsistent Quality and Performance - Batch-to-batch variability is common when using plant lectins. The quality of the products depends on the methodology of isolation of the plant material, and on the quality of the starting plant material itself.
• Incompatibility with recombinant production methods - Like many plant and eukaryotic proteins, plant lectins are not amenable to recombinant expression in E. coli. Therefore, expression often results in low yields or insoluble protein products. This is further complicated by the multiple subunits and post-translational modifications that eukaryotic lectins carry.
• Production is not readily scalable - The incompatibility of plant and eukaryotic lectins with recombinant production methods limits the scalability of their production, meaning their application is limited to small scale applications.

RPL products developed by Dublin City University and GlycoSeLect Ltd exhibit specificity and high affinity for a range of glycan epitopes displaying terminal galactose and mannose (including high mannose structures). All of our RPL products have a poly-histidine tag enabling simple detection using standard conjugated anti-his detection.

• Simple and Selective Production - Because of their origin, prokaryotic lectins are more amenable to recombinant expression in high levels in E. coli. The genetically incorporated histidine tag enables one-step, high-purification purification. In addition, using Stirred Tank Reactor systems, RPL expression is readily scalable. The scalability and plus the ease of purification, makes using RPL a cost-effective solution for your glycobiology.
• Consistent High Quality and Purity - Recombinant expression and selective affinity purification of RPL means products are consistently high in quality, purity, and performance. Plant lectins, on the other hand, often suffer from large batch-to-batch variation.
• Superior Performance - RPL exhibit greater selectivity and higher affinity for glycosylated targets than their plant and eukaryotic lectin equivalents. RPL exhibit excellent stability enabling RPL bioaffinity matrices to be reused many times. Because of their prokaryotic origins, they contain no disulphide bonds, and therefore can be used in the presence of denaturing and reducing conditions.

Lectin Engineering: Both RPL-Gal2 (left) and RPL-Gal1 (right) were derived from the alpha-galactophilic lectin RPL-aGal through the use of site specific mutagenesis. RPL-Gal2 exhibits significantly enhanced affinity for terminal alpha-linked galactose than either the parental RPL-aGal or the comparable plant lectin GSL-1 (approx 5 fold). RPL-Gal1 exhibits high affinity binding to terminal beta-1,4-linked galactose, to which the parental RPL-aGal doesn't bind, significantly greater than that of the comparable plant lectin ECL (approx 10 fold).