Preparation of Low Cost Affinity Chromatographic Matrix and Its Application in Purification of a Lectin Isolated from Mulberry Seeds
International Journal of Biomedical Materials Research
Volume 6, Issue 2, June 2018, Pages: 50-56
Received: Jun. 22, 2018; Accepted: Jul. 17, 2018; Published: Aug. 30, 2018
Views 1015      Downloads 98
Mosammat Jesmin Sultana, Department of Materials Science and Engineering, University of Rajshahi, Rajshahi, Bangladesh
Mohammad Taufiq Alam, Department of Applied Chemistry and Chemical Engineering, University of Rajshahi, Rajshahi, Bangladesh
Fazle Rabbi Shakil Ahmed, Department of Pharmacy, Khwaja Yunus Ali University, Sirajgonj, Bangladesh
Article Tools
Follow on us
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.
Immobilization, Affinity Matrix, Affinity Chromatography, Epichlorohydrin, Lectin, Hemagglutination
To cite this article
Mosammat Jesmin Sultana, Mohammad Taufiq Alam, Fazle Rabbi Shakil Ahmed, Preparation of Low Cost Affinity Chromatographic Matrix and Its Application in Purification of a Lectin Isolated from Mulberry Seeds, International Journal of Biomedical Materials Research. Vol. 6, No. 2, 2018, pp. 50-56. doi: 10.11648/j.ijbmr.20180602.14
Copyright © 2018 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License ( which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
D. K. Roper, E. N. Lightfoot, Separation of biomolecules using adsorptive membranes, J. Chromatogr. A 702 (1995) 3–26.
H. Abou-Rebyeh, F. Korber, K. Schubert-Rehberg, J. Reusch, D. Josic, Carrier membrane as a stationary phase for affinity chromatography and kinetic studies of membrane-bound enzymes, J. Chromatogr. A 566 (1991) 341–350.
R. Woker, B. Champluvier, M. R. Kula, Purification of S-oxynitrilase from Sorghum bicolor by immobilized metal ion affinity chromatography on different carrier materials, J. Chromatogr. B 584 (1992) 85–92.
C. Charcosset, Purification of proteins by membrane chromatography, J. Chem. Technol. Biotechnol. 71 (2) (1998) 95–110.
X. F. Zeng, E. Ruckenstein, Macroporous chitin affinity membranes for wheat germ agglutinin purification from wheat germ, J. Membr. Sci. 156 (1999) 97–107.
D. Josic, J. Reusch, K. Loster, O. Raum, W. Reutter, High performance membrane chromatography of serum and plasma membrane proteins, J. Chromatogr. 590 (1992) 59–76.
T. B. Tennikova, B. G. Belenkii, F. Svec, High-performance membrane chromatography a novel method of protein separation, J. Liq. Chromatogr. 13 (1990) 63–70.
M. Unarska, P. A. Davies, M. P. Esnouf, B. J. Bellhouse, Comparative-study of reaction-kinetics in membrane and agarose bead affinity systems, J. Chromatogr. A 519 (1991) 53–67.
N. Kubota, Y. Nakagawa, Y. Eguchi, Recovery of serum proteins using cellulosic affinity membrane modified by immobilization of Cu2+ ion, J. Appl. Polym. Sci. 62 (1996) 1153–1160.
S. Sternberg, Therapeutic plasmapheresis membrane device, Proc. Am. Chem. Soc. Div. Polym. Mater. Sci. Eng. 61 (1989) 665–669.
A. Malakian, M. Golebiowska, J. Bellefeuille, Purification of monoclonal and polyclonal IgG with affinity membrane matrix coupled with proteins A and G, Am. Lab. 40 (1993) 40.
Sharon J, Kabat EA, Morrison SL, Studies on mouse hybridomas secreting IgM or IgA antibodies to alpha(1 to 6)-linked dextran, Mol immunol. (1981); 18(9); 831-846.
N. Abser, T. Yeasmin, M. Salim Raza, S. Kumar Sarkar, F. Arisak, Single step purification, characterization and N-terminal sequences of a mannose specific lectin from mulberry seeds. Protein J. 24(2005)369–377.
Zhengyi Wu, Zhe-Kun Zhou , Michael G. Gilbert. "Ficus elastica". Flora of China. Missouri Botanical Garden, St. Louis, MO & Harvard University Herbaria, Cambridge, MA. Retrieved 29 August 2013.
Burgess KS, Morgan M, De Verno LL, Husband CB, Asymmetrical introgression between two Morus species (M. alba, M. rubra) that differ in abundance. Mol. Ecol. 14(2005) 3471-3483.
M. A. Hossain, S. M. R. Islam, N. Absar, Purification and characterization of lectins from mulberry seeds (Morus alba L.), Pakistan J Biol Sci. 7(2004) 1808-1813.
E. Ruckenstein, W. Guo, Crosslinked mercerized cellulose membranes and their application to membrane affinity chromatography, J. Membr. Sci. 187 (2001)277-286.
W. Guo, E. Ruckenstein, Crosslinked mercerized cellulose membranes for the affinity chromatography of papain inhibitors, J. Membr. Sci. 197(2002)53-62.
W. Guo, E. Ruckenstein, Separation and purification of horseradish peroxidase by membrane affinity chromatography, J. Membr. Sci. 211(2003)101-111.
U. K. Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227(5259) (1970)680-685.
S. R. Kabir, A. Hossen, M. A. Zubair, M. J. Alom, F. Islam, A. Hossain, Y. Kimu, A new lectin from the tuberous rhizome of Kaempferia rotunda: Isolation, characterization, antibacterial and antiproliferative activities, Protein Pept. Lett. 18(11)(2011)1140-1149.
J. Chen, B. Liu, N. Ji, J. Zhou, H, J. Bian, C. Y. Li, F. Chen, J. K. Bao, A novel sialic acid-specific lectin from Phaseolus coccineus seeds with potent antineoplastic and antifungal activities, Phytomed. 16(4) (2009)352-360.
A. K. M. Asaduzzaman, I. Hasan, A. Chakrabortty, Mst. S. Zaman, S. S. Islam, F. R. S. Ahmed, K. M. A. Kabir, M. Nurujjaman, M. B. Uddin, M. T. Alam, R. K. Shaha, S. R. Kabir, Moringa oleifera seed lectin inhibits Ehrlich ascites carcinoma cell growth by inducing apoptosis through the regulation of Bak and NF-κB gene expression, International Journal of Biological Macromolecules, 107 (2018) 1936-1944.
Une S, Nonaka K, Akiyama J, Lectin Isolated from Japanese Red Sword Beans (Canavalia gladiata) as a Potential Cancer Chemopreventive Agent, Journal of food science, 83(3)(2018) 837-843.
Science Publishing Group
1 Rockefeller Plaza,
10th and 11th Floors,
New York, NY 10020
Tel: (001)347-983-5186