See also Glycomics under Proteomics. 

See also Separation of Glycan variants in antibody purification

Lectin based affinity matrixes have been in use for many years and are used to capture and purifiy glycosylated proteins. Concanavalin A is one of the favored glycoprotein affinity lectins because it binds to a commonly occurring sugar structure in glycoproteins; alpha linked mannose. 

To enrich proteins carrying a specific glycan structure, lectin affinity chromatography has been used, for example using the Aleuria aurantia lectin which binds fucose containing glycans (Drake, “A lectin affinity workflow targeting glycosite-specific, cancer related carbohydrate structures in trypsin-digested human plasma. Anal Biochem. 408: 71-85 (2011). 

Allison (US14/215370) discloses a method or purifying recombinant polypeptides such as antibodies which have been produced in yeast or filamentous fungal cells using chromatography techniques such as Affintiy-Mixed Mode-HIC which includes lectin binding assays (using a lectin that binds to the glycosylated impurities such as a glycovariant of the antibody) to monitor for glycosylated impurities. 

Haberger (US2011/0117601 & WO2009/027041) discloses a method for the production of a glycosylated hterologous polypeptide comprising the steps of obtaining a sample form a crude fermentation broth, incubating the same with magnetic affinity beads to which Protein A, G, L or lectin is bound, releasing glycans from the immobilized glycosylated polypeptides without the polypeptide from being released, purifying the glycan and measuring a glycosylation profile as by mass spetrometry by comparing the glycosylation profile with a desired glycosylation profile of the recombinant glycosylated polypetide and modifying the culture conditions in accordance to the glycosylation profile. 

Regnier (US8,568,993) discloses isolation of glycosylated peptides or proteins using lectin or antibody affinity chromatography. The eluate containing fucosylated peptides was collected for HPLC fractionation and MALDI-TOF analysis. 

Schnorr (WO2004/090549) dicloses provising a host cell which secrets a glycosylated polypeptide of interest and contacting a sample comprising the glycosylated polypeptide of interest with an immobilized lectin capable of binding to the glycosylated polypeptide of interest, under conditions where binding capacity of the immobilized lectin is at least 10 ng polypeptide/200 micol compartment volume. 

Sethi (“using single lectins to enrich glycoproteins in conditioned media” Unit 24.6 from Current Protocols in Protein Science (2007) discloses a protocol for a single lectin based affinity chromtography which uses a single agarose bound lecitn to capture and enrich glycoproteins from serum free conditioned media. 

Xu (Mol Biotechnol (2010) 45: 248-256) discloses using Lectin ECA) which has a primary reactivityy to Gal beta1, 4 GlcNAc into a cell based and ELISA based binding assay to measure the degree of Galbeta1,4 GlcNAc termianl exposure on Human FVIII. The level of Galbeta1,4 exposure is inversely corrected to the protein/FVIII sialyation level so that the level of beta1,4 galactose terminal exposure indirectly reflects the sialyation status of the glycoprotein.

Applications

Diagnosis of Cancer based on glycosylation profile: See glycomics under “proteomics”

Modulation of Cell Culture:

Laine disclsoes methods to modulate and grow stem cells by contacting stem cells with a binder recognizing terminal glycan structure of stem cells. In one embodiment, a lectin immobilized on a surface is used as a capture agent to bind the particular glycan structure.