Enhanced Glycoselective Bioaffinity Proteins
AMSBIO has announced the introduction of a range of Recombinant Prokaryotic Lectins (RPLs). RPLs are enhanced glycoselective bioaffinity proteins that enable efficient detection, analysis and selective isolation of glycosylated biomolecules.
RPLs offer many advantages over traditional plant and eukaryotic lectins. They also offer many advantages over current glycoanalytical and chromatographic methodologies that are not glycoselective or require prior release of glycans which results in the sample destruction.
Recombinant production methods present opportunities to manipulate RPL glycan binding properties through the use of site specific mutagenesis strategies. Lectins can be engineered to enhance their affinity for a given glycan moiety or to alter their specificity for alternative glycan moieties. This can allow RPL properties to be tailored for specific targets or to enhance their performance for specific applications.
RPLs exhibit excellent stability enabling RPL bioaffinity matrices to be reused many times. Because of their prokaryotic origins RPLs lack the presence of disulphide bonds. This allows RPLs to be used in the presence of reducing agents for applications under denaturing conditions.
In standard ELLA formats RPLs enable significantly greater sensitivity of detection at lower lectin concentrations than that required for plant lectins. RPLs exhibit greater selectivity and higher affinity for glycosylated targets than comparable plant and eukaryotic lectins. RPLs are expressed with genetically incorporated poly-histidine tags that enable simple one step and high capacity purification of products via immobilized metal affinity chromatography (IMAC).
Recombinant expression and selective affinity purification of RPLs results in the production of products of consistent high quality, purity and performance. This is in contrast to plant lectins for which variation in performance (from batch to batch and from one supplier to another) has traditionally been reported.
RPLs expression is readily scalable using Stirred Tank Reactor (STR) systems. This combined with the high capacity and scalability of IMAC purification enables cost effective production of RPLs in sufficient quantities required for large scale applications in the separation and purification of glycosylated biomolecules.