title: Development of monoclonal antibody-based sandwich ELISA for detection of dextran.
abstract: Dextran as anti-nutritional factor is usually a result of bacteria activity and has associated serial problems during the process stream in the sugar industry and in medical therapy. A sensitive method is expected to detect dextran quantitatively. Here we generated four monoclonal antibodies (MAbs) against dextran using dextran T40 conjugated with bovine serum albumin (BSA) as immunogen in our lab following hybridoma protocol. Through pairwise, an MAb named D24 was determined to be conjugated with horseradish peroxidase (HRP) and was used in the establishment of a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) method for determination of dextran, in which MAb D9 was chosen as a capture antibody. The detection limit and working scope of the developed sandwich ELISA method were 3.9 ng/mL and 7.8-500 ng/mL with a correlation coefficient of 0.9909. In addition, the cross-reaction assay demonstrated that the method possessed high specificity with no significant cross-reaction with dextran-related substances, and the recovery rate ranged from 96.35 to 102.00%, with coefficient of variation ranging from 1.58 to 6.94%. These results indicated that we developed a detection system of MAb-based sandwich ELISA to measure dextran and this system should be a potential tool to determine dextran levels.
authors.affiliation: 1 Cancer Research Center, Medical College, Xiamen University , Xiangan South Road, Xiamen, Fujian, China .
authors.forename: Sheng-Yu
authors.initials: SY
authors.lastname: Wang
authors.forename: Zhe
authors.initials: Z
authors.lastname: Li
authors.forename: Xian-Jiang
authors.initials: XJ
authors.lastname: Wang
authors.forename: Sha
authors.initials: S
authors.lastname: Lv
authors.forename: Yun
authors.initials: Y
authors.lastname: Yang
authors.forename: Lian-Qiang
authors.initials: LQ
authors.lastname: Zeng
authors.forename: Fang-Hong
authors.initials: FH
authors.lastname: Luo
authors.forename: Jiang-Hua
authors.initials: JH
authors.lastname: Yan
authors.forename: Da-Feng
authors.initials: DF
authors.lastname: Liang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D003429:Cross Reactions / Q000276:immunology; D003911:Dextrans / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006735:Horseradish Peroxidase / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D057230:Limit of Detection; D051379:Mice; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D003911:Dextrans; D006735:Horseradish Peroxidase