Improving immunobinding using oriented immobilization of an oxidized antibody
- Title
- Improving immunobinding using oriented immobilization of an oxidized antibody
- Author
- 이은규
- Keywords
- magnetic particles; immunobinding; mouse IgG; oriented immobilization; ligand antibody; surface modification
- Issue Date
- 2007-08
- Publisher
- ELSEVIER SCIENCE BV
- Citation
- JOURNAL OF CHROMATOGRAPHY A, v. 1161, No. 1-2, Page. 9-14
- Abstract
- Recent technical advances in biorecognition engineering and microparticle fabrication enabled us to develop a single-step purification process using magnetic particles (MPs). The process is simple, efficacious, easy to automate, and economical. The method immobilizes the ligand molecule in a particular orientation on commercial MPs that have surface carboxyl groups. Mouse IgG and anti-mouse IgG antibody were the model capture and ligand molecules for this study. The immunobinding efficacy of anti-mouse IgG antibody using "oriented immobilization" was compared with the efficacy of a conventional amine-coupling system that results in random orientation and of another standard method, the biotin-streptavidin system. The oriented immobilization was accomplished by oxidizing the sugar moiety in the CH2 domain of the antibody's Fc and covalently conjugating the moiety to the hydrazine-coated MP. The specific binding affinity of the oriented immobilization process was about 2.5 times that of the amine-coupling system, and selectivity from a binary mixture was about 2 times greater for the oriented immobilization method. Results were nearly identical for the biotin-streptavidin system and the oriented immobilization system, matching the calculated binding stoichiometry between mouse IgG and anti-mouse IgG antibody. The binding improvement over the amine-coupling system shown by assay was confirmed by a separate surface plasmon resonance experiment. In summary, the oriented immobilization method was as effective as the streptavidin-biotin system, yet simpler and cost-effective. (c) 2007 Elsevier B.V. All rights reserved.
- URI
- https://www.sciencedirect.com/science/article/pii/S0021967307008679https://repository.hanyang.ac.kr/handle/20.500.11754/106822
- ISSN
- 0021-9673; 1873-3778
- DOI
- 10.1016/j.chroma.2007.05.023
- Appears in Collections:
- COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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