title: PEGylation, detection and chromatographic purification of site-specific PEGylated CD133-Biotin antibody in route to stem cell separation.
abstract: Recovery and purification of stem cells are determining steps in order to obtain the purity and viability required for transplantation. In this context, immunochemical techniques have been widely preferred due to their high selectivity. CD133, a glycoprotein expressed by stem cells, is a well-used marker for isolation of neural stem cells. Transplantation of neural stem cells into patients can promote neural growth and improve neuronal functions. In this study, a new method for site-specific PEGylation of CD133-Biotin antibody is performed through streptavidin-biotin conjugation. Purification was carried out by ion-exchange chromatography. The characterization of the single PEGylated CD133-Biotin antibody was confirmed using electrophoresis with silver staining and I(2)-BaCl(2) for PEG detection. Moreover, online PEG quantification directly after the chromatographic step was conducted (in each fraction) to detect exact elution times of PEG. In conclusion, the novel CD133-Biotin antibody PEGylation strategy conducted in this study could be used as a process step in route to neural stem cell recovery and purification via the modification of existing techniques such as aqueous two phase systems, PEGylated affinity columns or fluidized chromatography.
authors.affiliation: Centro de Biotecnología-FEMSA, Tecnológico de Monterrey, Campus Monterrey, Ave. Eugenio Garza Sada 2501 Sur, Monterrey, NL 64849, Mexico.
authors.forename: Mirna
authors.initials: M
authors.lastname: González-González
authors.forename: Karla
authors.initials: K
authors.lastname: Mayolo-Deloisa
authors.forename: Marco
authors.initials: M
authors.lastname: Rito-Palomares
mesh_terms: D000071916:AC133 Antigen; D000906:Antibodies / Q000737:chemistry* / Q000378:metabolism; D015703:Antigens, CD / Q000737:chemistry* / Q000276:immunology; D017609:Barium Compounds; D001710:Biotin / Q000737:chemistry* / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D002712:Chlorides; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology; D010455:Peptides / Q000737:chemistry* / Q000276:immunology; D011092:Polyethylene Glycols / Q000737:chemistry*; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D000906:Antibodies; D015703:Antigens, CD; D017609:Barium Compounds; D002712:Chlorides; D006023:Glycoproteins; D010455:Peptides; C024986:barium chloride; D011092:Polyethylene Glycols; D001710:Biotin; D019809:Streptavidin