I am trying to purify glycosylated fibronectin from serum samples. Unfortunately, there is no specific antibody against glycosylated fibronectin. So, I want to use lectin-based affinity chromatography to isolate total N-glycans from serum first, and then do the western blot using plasma fibronectin antibody. Is this approach correct? And which lectin should I use, WGA or Con-A?
Sarojini: It depends on what degree of purity you want to achieve for the glycosylated fibronectin. As secreted glycoproteins contain complex carbohydrate, they bind poorly on Con A. So, WGA maybe a good choice, in conjunction with other traditional steps such as ion exchange, size exclusion etc. Good luck.
Ke-Wei Zhao Thank you for your suggestion. I was going through literature and found an article, where they have shown maximum binding of glycosylated fibronectin with ConA and least binding with WGA. Article Lectins as probes for assessing the accessibility of N‐linke...
Please let me know your thoughts on this.
Sarojini: Well, experiment (wet science) always overrides theories. Use that article as a guide for designing your own one. Good luck.
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