title: Optimal activation of carboxyl-superparamagnetic iron oxide nanoparticles bioconjugated with antibody using orthogonal array design.
abstract: This study aims to bioconjugate anti-EMMPRIN monoclonal antibody on the surface of carboxyl-SPIO nanoparticles and to optimize the activated conditions of bioconjugation. Anti-EMMPRIN monoclonal antibody bioconjugated carboxyl-SPIO nanoparticles were performed through a coupling strategy of EDC and sulfo-NHS. The procedure was comprised of two steps by activation of carboxyl-SPIO nanoparticles and conjugation with monoclonal antibody. The optimal activated parameters of bioconjugation were evaluated by single factor design and orthogonal array design. SDS-PAGE analysis and Bradford assay was used for testing and verifying the efficiency of activated conditions obtained from orthogonal array. The results show that pH value, temperature and reaction time were important factors that influence bioconjugated efficiency. The activated parameters with pH value 6.2, temperature 25 degrees C and reaction time 30 min were obviously optimal for activation of carboxyl-SPIO nanoparticles and conjugation with monoclonal EMMPEIN antibody. This coupling strategy for anti-EMMPRIN mAb bioconjugated on SPIO nanoparticles was efficient, and may be further applied in the fields of medical or biological practices.
authors.affiliation: School of Public Health, Southeast University, Nanjing 210009, Jiangsu, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Liu
authors.forename: Xiaoqang
authors.initials: X
authors.lastname: Zhang
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yuepu
authors.initials: Y
authors.lastname: Pu
authors.forename: Lihong
authors.initials: L
authors.lastname: Yin
authors.forename: Meng
authors.initials: M
authors.lastname: Tang
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005290:Ferric Compounds / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008280:Magnetics; D053768:Metal Nanoparticles / Q000737:chemistry*; D013696:Temperature
chemical_list: D000906:Antibodies; D005290:Ferric Compounds; C000499:ferric oxide