Lectin-binding assay by polyethylene glycol 8000

Abidi, Fatima E. ; Bishayee, Subal ; Bachhawat, Bimal K. ; Bhadra, Ranjan (1987) Lectin-binding assay by polyethylene glycol 8000 Analytical Biochemistry, 166 (2). pp. 257-266. ISSN 0003-2697

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Official URL: http://linkinghub.elsevier.com/retrieve/pii/000326...

Related URL: http://dx.doi.org/10.1016/0003-2697(87)90573-2


A quantitative lectin-binding assay using a precipitation technique and polyethylene glycol 8000 (PEG) as a precipitating agent has been described. Carcinoscorpin, a sialic acid-binding lectin isolated from the hemolymph of Indian horseshoe crab, Carcinoscorpius rotunda cauda, and iodinated fetuin, a sialoglycoprotein, were appropriately incubated as the components of the binding assay. The specific interaction between these two components developed the lectin-glycoprotein-bound complex. This was subsequently precipitated by the addition of PEG together with a coprecipitant γ-globulin. Radioactivity of the precipitated bound complex was estimated to quantify the binding. The formation of the bound complex was effectively inhibited by a specific sialodisaccharide, O-(N-acetylneuraminyl)-(2 → 6)-2-acetamido-2-deoxygalactitol, implying the specific interaction for such precipitation. The probable effect of PEG was to stabilize the bound complex, precipitating it along with added γ-globulin. This was further evident from the prevention of dissociation of the bound complex and increased binding of glycoprotein to the imobilized lectin in the presence of PEG. The assay was also applicable to other sialoglycoproteins such as α1-acid glycoprotein and human chorionic gonadotropin. Moreover, the method yielded a saturation plateau with a characteristic hyperbolic binding curve. The assay was simple, quick, safe, economic, and highly sensitive.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:Lectin (carcinoscorpin); Binding Assay; Polyethylene Glycol 8000; Precipitation; Stabilization
ID Code:2281
Deposited On:07 Oct 2010 11:38
Last Modified:12 May 2011 11:34

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