Main Article Content


A.B. Nurmagambetova

National Center for Biotechnology, 13/5 Korgalzhyn road, Astana, Z05K8D5, Kazakhstan

D.G. Akhmetova

Republican Diagnostic Center, HighVill B-1, 3 Akhmet Baitursynov, Astana, 020000, Kazakhstan

B.B. Khassenov

National Center for Biotechnology, 13/5 Korgalzhyn road, Astana, Z05K8D5, Kazakhstan

K.K. Baltin

National Center for Biotechnology, 13/5 Korgalzhyn road, Astana, Z05K8D5, Kazakhstan


Recombinant protein A is widely used in biotechnology for the purification of immunoglobulin G antibodies. In this study, truncated protein A (protein A (5), encoding only five IgG binding domains (E, D, A, B, and C) and lacking signal sequence S and cell-wall anchoring region X M, was cloned into pMBP his parallel 2 and expressed in BL21 (DE3) cells. The gene encoding protein A was inserted downstream of the maltose binding protein (MBP) encoding malE gene. MBP is often fused with other proteins to improve their solubility, enhance stability, and increase the final yield. Proteins expressed using this system can be found in the soluble fraction, which is likely due to the solubilizing properties of MBP. A strain producing recombinant protein A (5) fused with MBP was obtained. The recombinant protein was purified and concentrated by gradient affinity chromatography. The weight of the protein fused with MBP was approximately 77.3 kDa, and without MBP was approximately 37 kDa. The IgG binding specificity of the recombinant protein was assessed using the agar gel immunodiffusion test with the target protein, bull, mouse, rat, and rabbit IgGs. The antigen antibody interaction showed a pattern not character to that described previously.


protein A, cloning, expression, purification, spa, SpA

Article Details


Boström T., Nilvebrant J., Hober S. Purification systems based on bacterial surface proteins. Protein Purification., 2012.

Sidorin E.V., Solov’eva T.F. IgG Binding Proteins of Bacteria. Biochemistry, 2011, vol. 76, no. 3, pp. 363-378.

Uhlen M., Guss B., Nilsson B., Gatenbeck S., Philipson L., Lindberg M. Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications. J Biol Chem., 1984, vol. 259, no.3, pp. 1695-1702.

Olsson A., Eliasson M., Guss B., Nilsson B., Hellman U., Lindberg M., Uhlen M. Structure and evolution of the repetitive gene encoding streptococcal protein G. European Journal of Biochemistry, 1987, vol. 168, no. 2, pp. 319-324.

Akerström B., Björck L. Protein L: an immunoglobulin light chain-binding bacterial protein. Characterization of binding and physicochemical properties. Journal of Biological Chemistry, 1989, vol. 264, no. 33, pp. 19740-19746.

Bjorck L. Protein L. A novel bacterial cell wall protein with affinity for Ig L chains. J Immunol, 1988, vol. 140, no. 4, pp. 1194-1197.

Iangone J. J. Protein A of Staphylococcus aureus and Related lrnmunoglobulin Receptors Produced by Streptococci and Pneumonococci. Advances in immunology, 1982, vol. 32, pp.157-252.

Jendeberg L., Nilsson P., Larsson A., Denker P., Uhlen M., Nilsson B., Nygren P. A. Engineering of Fc 1 and Fc 3 from human immunoglobulin G to analyse subclass specificity for staphylococcal protein A. Jourmal of Immunological Methods, 1997, vol. 201, no. 1, pp. 25-34

Ljungberg U. K., Jansson B., Niss U., Nilsson R., Sandberg B.E., Nilsson B. The interaction between different domains of staphylococcal protein A and human polyclonal IgG, IgA, IgM and F (ab') 2: separation of affinity from specificity. Molecular immunology, 1993, vol. 30, no. 14, pp. 1279-1285.

Goward C. R., Murphy J. P., Atkinson T., Barstow D. A. Expression and purification of a truncated recombinant streptococcal protein G. Biochemical Journal, 1990, vol. 267, no.1, pp.171-177.

Abrahmsen L., Moks T. Nilsson B. Hellman U. Uhlen M. Analysis of signals for secretion in the staphylococcal protein A gene. EMBO J 4, 1985, vol. 4, no.13B, pp.3901-3906.

Hollingshead S. K., Fischetti V. A., Scott J. R. Complete nucleotide sequence of type 6 M protein of the group A Streptococcus. Repetitive structure and membrane anchor. Journal of Biological Chemistry, 1986, vol. 261. no. 4, pp.1677-1686.

Boyle M. D. P. The type I bacterial immunoglobulin-binding protein: staphylococcal protein A.Bacterial immunoglobulin-binding proteins, 1990, vol. 1, pp.17-28.

Rigi G., Beyranvand P., Ghaedmohammadi S., Heidarpanah S., Noghabi K. A., Ahmadian G. Comparison of the extracellular full-length and truncated recombinant protein A production in Escherichia coli BL21 (DE3). Journal of Paramedical Sciences, 2015,vol. 3.

Eliasson M., Olsson A., Palmcrantz E., Wiberg K., Inganäs M., Guss B., Uhlen M. Chimeric IgG-binding receptors engineered from staphylococcal protein A and streptococcal protein G.Journal of Biological Chemistry,1988, vol. 263. no. 9, pp. 4323-4327.

Hober S., Nord K., Linhult M. Protein A chromatography for antibody purification. Journal of Chromatography B, 2007, vol. 848, no. 1, pp. 40-47.

Sheffield P., Garrard S., Derewenda Z. Overcoming expression and purification problems of RhoGDI using a family of “parallel” expression vectors. Protein expression and purification, 1999, vol.15, no.1, pp. 34-39.

Khassenov B.B., Sultankulov B.M., Kozhahmetov S.S., Akhmetov S.B., Ramanklov E.M., Mukanov K.K. Poluchenie rekombinantnogo antigena lejkoza virusa krupnogo rogatogo skota p24.Biotechnology. Theory and Practice, 2011, no. 2, pp.50-56.

Abeldenov S.K. Cloning, expression and purification of recombinant analog of Taq DNA polymerase. Biotechnology. Theory and Practice, 2014, no.1, pp.12-16.

Mussakhmetov A., Nurmagambetova A., Abeldenov S., Khassenov B. Purification of recombinant Pfu DNA polymerase by double step affinity chromatography. Biotechnology. Theory and Practice, 2014, no.2, pp. 42-47.

Abeldenov S., Kirillov S., Nurmagambetova A., Kiribayeva A., Silayev D., Khassenov B. Expression and purification and biochemical characterization of recombinant phosphohydrolaseappa in Esherichia coli. Biotechnology. Theory and Practice, 2014, no.3, pp. 61-65.

Ouchterlony, Ö. Antigen‐antibody reactions in gels. APMIS, 1953, vol.32, no.2, pp. 231-240.

Justiz-Vaillant A. A., Akpaka P. E., McFarlane-Anderson N., Smikle M. F. Comparison of techniques of detecting immunoglobulin-binding protein reactivity to immunoglobulin produced by different avian and mammalian species. West Indian Medical Journal, 2013, vol.62, no.1, pp.12-20.

Richman D. D., Cleveland P. H., Oxman M. N., Johnson K. M. The binding of staphylococcal protein A by the sera of different animal species. The Journal of Immunology, 1982, vol. 128, no. 5, pp. 2300-2305.

Frank M. B. Antibody Binding to Protein A and Protein G beads. Molecular Biology Protocols. Oklahoma City, 1997.