Partitioning and recovery of Canavalia brasiliensis lectin by aqueous two-phase systems using design of experiments methodology

The performance of aqueous two-phase systems (ATPS) composed by poly(ethylene glycol) (PEG)- and different salts (sodium citrate, potassium phosphate, sodium sulfate and ammonium sulfate) has been evaluated and compared for the selective isolation of Canavalia brasilienses lectin (ConBr) from crude extracts of C. brasilienses seeds. Among the systems studied, PEG-phosphate was the most efficient for the purification of ConBr. A central composite design was applied to study the effects of different factors such as, PEG concentration, phosphate buffer concentration, sodium chloride concentration and pH on ConBr extraction and to optimize its isolation from a plant extract. ATPS comprising PEG 600–phosphate buffer provided a means for the recovery of proteins from crude extracts of C. brasilienses seeds. The best conditions of purification were achieved using an ATPS composed by 16.5% (w/w) PEG 600, 15.0% (w/w) phosphate pH 7.5, 4.5% (w/w) NaCl. The maximum percentage yield of protein extracted was about 100% with a final purity of 73.04%.

[1]  M. Kula,et al.  Investigation of mathematical methods for efficient optimisation of aqueous two-phase extraction. , 2000, Journal of chromatography. B, Biomedical sciences and applications.

[2]  B. Cavada,et al.  Characteristics of the histamine release from hamster cheek pouch mast cells stimulated by lectins from Brazilian beans and concanavalin A , 1996, Inflammation Research.

[3]  Ana M Azevedo,et al.  Application of central composite design to the optimisation of aqueous two-phase extraction of human antibodies. , 2007, Journal of chromatography. A.

[4]  W. Legnani.  Mistletoe in Conventional Oncological Practice: Exemplary Cases , 2008, Integrative cancer therapies.

[5]  M. Eberlin,et al.  Cloud point extraction applied to casein proteins of cow milk and their identification by mass spectrometry. , 2007, Analytica chimica acta.

[6]  D. Shaw,et al.  Introduction to colloid and surface chemistry , 1970 .

[7]  K. Berggren,et al.  Effects of salts and the surface hydrophobicity of proteins on partitioning in aqueous two-phase systems containing thermoseparating ethylene oxide-propylene oxide copolymers , 1995 .

[8]  B. Cavada,et al.  Lectin-induced nitric oxide production. , 1999, Cellular immunology.

[9]  Ana M Azevedo,et al.  Chromatography-free recovery of biopharmaceuticals through aqueous two-phase processing. , 2009, Trends in biotechnology.

[10]  R. H. Khan,et al.  Partially folded intermediate state of concanavalin A retains its carbohydrate specificity. , 2005, Biochemical and biophysical research communications.

[11]  V. Kher,et al.  Reduction of Hepatitis C Virus Using Lectin Affinity Plasmapheresis in Dialysis Patients , 2009, Blood Purification.

[12]  B. Cavada,et al.  Revisiting proteus: do minor changes in lectin structure matter in biological activity? Lessons from and potential biotechnological uses of the Diocleinae subtribe lectins. , 2001, Current protein & peptide science.

[13]  B. Cavada,et al.  In vivo lymphocyte activation and apoptosis by lectins of the Diocleinae subtribe. , 2001, Memorias do Instituto Oswaldo Cruz.

[14]  D. Prazeres,et al.  Plasmid purification by hydrophobic interaction chromatography using sodium citrate in the mobile phase , 2009 .

[15]  M. Aires-Barros,et al.  Downstream processing of human antibodies integrating an extraction capture step and cation exchange chromatography. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[16]  N. Sharon,et al.  History of lectins: from hemagglutinins to biological recognition molecules. , 2004, Glycobiology.

[17]  P. Rougé,et al.  Plant Lectins: A Composite of Several Distinct Families of Structurally and Evolutionary Related Proteins with Diverse Biological Roles , 1998 .

[18]  G. Hall.  Methods of Testing Protein Functionality , 2011 .

[19]  Marco Rito-Palomares,et al.  Practical application of aqueous two-phase partition to process development for the recovery of biological products. , 2004, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[20]  M. Kaster,et al.  Antidepressant‐like effect of lectin from Canavalia brasiliensis (ConBr) administered centrally in mice , 2006, Pharmacology Biochemistry and Behavior.

[21]  M. del-Val,et al.  Biphasic aqueous media containing polyethylene glycol for the enzymatic synthesis of oligosaccharides from lactose , 2003 .

[22]  Been-Huang Chiang,et al.  Partitioning and purification of lysozyme from chicken egg white using aqueous two-phase system , 2006 .

[23]  Ana M. Azevedo,et al.  Partitioning of human antibodies in polyethylene glycol–sodium citrate aqueous two-phase systems , 2009 .

[24]  M. Aires-Barros,et al.  Purification of human immunoglobulin G by thermoseparating aqueous two-phase systems. , 2008, Journal of chromatography. A.

[25]  A. Wlodawer,et al.  Structural studies of algal lectins with anti-HIV activity. , 2006, Acta biochimica Polonica.

[26]  Ana M Azevedo,et al.  Optimisation of aqueous two-phase extraction of human antibodies. , 2007, Journal of biotechnology.

[27]  Hans-Joachim Gabius,et al.  The sugar code: functional lectinomics. , 2002, Biochimica et biophysica acta.

[28]  M. Raida,et al.  Molecular characterization and crystallization of Diocleinae lectins. , 1999, Biochimica et biophysica acta.

[29]  J. Švitel,et al.  Lectinomics II. A highway to biomedical/clinical diagnostics. , 2009, Biotechnology advances.

[30]  E. Teixeira,et al.  Renal effects of the lectin from Canavalia brasiliensis seeds , 2001 .

[31]  J. Calvete,et al.  The crystal structure of Canavalia brasiliensis lectin suggests a correlation between its quaternary conformation and its distinct biological properties from Concanavalin A , 1997, FEBS letters.

[32]  B. Cavada,et al.  In vivo protective effect of the lectin from Canavalia brasiliensis on BALB/c mice infected by Leishmania amazonensis. , 1996, Acta tropica.