TY - JOUR
T1 - Boronate affinity chromatography of cells and biomacromolecules using cryogel matrices
AU - Srivastava, Akshay
AU - Shakya, Akhilesh Kumar
AU - Kumar, Ashok
N1 - Funding Information:
The authors acknowledge Department of Biotechnology (DBT), Ministry of Science & Technology, Govt of India . AS and AKS acknowledges senior research fellowships from University Grants Commission (UGC) and Council of Scientific and Industrial Research (CSIR), India, respectively.
PY - 2012/12/10
Y1 - 2012/12/10
N2 - Boronate affinity chromatography involves the interaction between cis-diol containing molecules and the hydroxyl group of boronate. Boronate affinity based cryogel chromatography matrices have been developed and the ligands were immobilized by two methods i.e., grafting of the boronate ligand on to the matrix and by copolymerization of monomer containing boronate with other co-monomer. The boronate grafted cryogel column was used to capture adherent and non-adherent cells and the captured cells were recovered at different fructose concentrations as an eluting agent, in chromatography mode. It was found that the adherent cells could be recovered at relatively higher fructose concentration (0.5. M) than non-adherent cells which could be recovered by using low fructose concentration (0.1. M). This might be due to the difference in the content of glycoprotein in adherent and non-adherent cells. In this way a new separation method can be devised for the fractionation of adherent and non-adherent cells. In another study, a copolymerized boronate cryogel column was developed for the separation of RNA from the bacterial crude extract without any pre-processing. RNA molecules were specifically retained in the cryogel column due to interaction between 2,3' diol group of ribose sugar in RNA and the hydroxyl group of boronate. The DNA molecules were passed through the column uninteracted due to absence of 2'-hydroxyl group. Later, bound RNA molecules were recovered from the boronate affinity cryogel column.
AB - Boronate affinity chromatography involves the interaction between cis-diol containing molecules and the hydroxyl group of boronate. Boronate affinity based cryogel chromatography matrices have been developed and the ligands were immobilized by two methods i.e., grafting of the boronate ligand on to the matrix and by copolymerization of monomer containing boronate with other co-monomer. The boronate grafted cryogel column was used to capture adherent and non-adherent cells and the captured cells were recovered at different fructose concentrations as an eluting agent, in chromatography mode. It was found that the adherent cells could be recovered at relatively higher fructose concentration (0.5. M) than non-adherent cells which could be recovered by using low fructose concentration (0.1. M). This might be due to the difference in the content of glycoprotein in adherent and non-adherent cells. In this way a new separation method can be devised for the fractionation of adherent and non-adherent cells. In another study, a copolymerized boronate cryogel column was developed for the separation of RNA from the bacterial crude extract without any pre-processing. RNA molecules were specifically retained in the cryogel column due to interaction between 2,3' diol group of ribose sugar in RNA and the hydroxyl group of boronate. The DNA molecules were passed through the column uninteracted due to absence of 2'-hydroxyl group. Later, bound RNA molecules were recovered from the boronate affinity cryogel column.
KW - Adherent cells
KW - Boronate affinity chromatography
KW - Cell separation
KW - Cryogel
KW - Non adherent cells
KW - RNA isolation
UR - http://www.scopus.com/inward/record.url?scp=84867194030&partnerID=8YFLogxK
U2 - 10.1016/j.enzmictec.2012.08.006
DO - 10.1016/j.enzmictec.2012.08.006
M3 - Article
C2 - 23040394
AN - SCOPUS:84867194030
SN - 0141-0229
VL - 51
SP - 373
EP - 381
JO - Enzyme and Microbial Technology
JF - Enzyme and Microbial Technology
IS - 6-7
ER -