Lectin Affinity Chromatography: An Efficient Method to Purify Horse IgG3.

This study presents an effective method for purifying a specific type of horse antibody, IgG3, using lectin affinity chromatography. This method relies on the unique binding interactions between lectins and their sugar-based counterparts in proteins.

Concepts and Basis of the Research

• The researchers are experimenting with affinity chromatography, a technique used to separate substances. The method is centered on the specific binding interaction established between a stationary substance (ligand) and its counterpart that moves and binds the ligand.
• This study focuses on a certain type of ligands called lectins. Lectins are molecules primarily found in plants that have the ability to bind specifically with different sugars. Because roughly one-third of all proteins are glycosylated, meaning sugars are added to them, many lectin-sugar pairs exist.
• The research uses these specific binding interactions between lectins and sugars to develop strategies for separating and understanding molecules. The methods also help investigate various biological phenomena such as tumor metastasis, intracellular communication, and inflammation.

Research Approach and Findings

• The researchers describe a procedure related to the separation of various horse antibody classes through affinity chromatography. The procedure relies primarily on the differences in the glycosylation patterns of these antibody classes.
• Using a Jacalin column (a specific lectin derived from the Artocarpus integrifolia), they have successfully segregated horse IgG3 from six other antibodies in horse serum in just one step.
• The significance of IgG3 lies in its medicinal application – it forms part of a therapeutic fraction created in equines for passive antibody therapy. Moreover, it can also serve as an indicator for patient hypersensitivity.
• Throughout the study, the team also determined the binding interaction constant between hypersensitive human immunoglobulin (huIgE) and the purified horse IgG3 (hoIgG3).