Abstract
Objective: Uniform sized agarose gel beads were prepared from agarose by emulsification technique. The prepared matrix has been attached with commercially available concanavalin A (conA) and finally it has applied to purify mulberry seed lectin. The present work shows a simple and inexpensive method for the preparation of an affinity matrix for purification of conA specific mulberry seed lectin. Method: Firstly, conA was immobilized on agarose gel beads and coupled to hexadiamine by using amino reactive bifunctional crosslinker (2,4,6-trichloro-1,3,5-trizine). Then it used as affinity matrix for the purification of mulberry seed lectin (designated as MSL). In purification protocol initially conA has been immobilized with agarose. Then MSL have been purified from the seeds of Morus alba L. Result: The agarose beads showed the best uniformity when 2-2.5% concentration of Tween 80 was used. The newly prepared affinity matrix agarose gel was able to purify MSL with the molecular weight of 22 kDa in a single step. The purified lectin strongly agglutinated with mice, chicken, bovine and human blood types A, B and O erythrocytes respectively. There was no activity found on goat erythrocytes. Conclusion: The present work shows a simple and inexpensive method for the preparation of an affinity chromatographic matrix. The prepared matrix was applied to purify mulberry seed lectin.
Highlights
In recent years, the membrane affinity chromatography of biomolecules had a fast development, because their chemical potential essential consistent and capable extend methods of separation and purification [1,2,3,4,5]
From the result mulberry seed lectin powerfully agglutinated with mice erythrocytes whereas no activity found on goat erythrocytes
This research article explained a simple and inexpensive method for the preparation of uniform sized agarose gel beads which was used as affinity chromatographic matrix
Summary
The membrane affinity chromatography of biomolecules had a fast development, because their chemical potential essential consistent and capable extend methods of separation and purification [1,2,3,4,5]. Several resources, such as nylon, polysulfone, chitosan, cellulose and cellulose derivatives [6,7,8,9,10,11] were used to prepare affinity chromatographic matrixes. For ligand coupling three immobilization steps were used for matrix preparation such as treatment with Epichlorohydrin, modification with 1, 6-Hexanediamine, activation with 2,4,6trichloro-1,3,5-trizine
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