High-throughput optimization of surfaces for antibody immobilization using metal complexes
- 作者：Ben W. Muir ; Michael C. Barden ; Simon P. Collett ; Alain-Dominique Gorse ; Raisa Monteiro ; Liqun Yang ; Nicole A. McDougall ; Sharon Gould ; N. Joe Maeji
- 关键词：High throughput ; Surface discovery ; ELISA ; Antibody immobilization ; Metal complexes
- 刊名：Analytical Biochemistry
- 出版年：2007
- 期刊代码：93_00032697
- 类别：imm
- 出版时间：1 April 2007
- 卷：363
- 期：1
- 页码：97-107
- 文件大小：860 K
摘要
Using a high-throughput surface discovery approach, we have generated a 1600-member library of metal-containing surfaces and screened them for antibody binding potential. The surface library assembly involved graft modification of argon plasma-treated polyvinylidenedifluoride (PVDF) membranes with alternating maleic anhydride-styrene copolymer followed by anhydride ring opening with a range of secondary amines and microarray contact printing of transition metal complexes. The microarrays of metal-containing surfaces were then tested for their antibody binding capacity by incubation with a biotinylated mouse antibody in a chemiluminescence assay. A total of 11 leads were identified from the first screen, constituting a “hit” rate of 0.7%. A smaller 135-member surface library was then synthesized and screened to optimize existing hits and generate additional leads. To demonstrate the applicability of these surfaces to other formats, high-binding surface leads were then transferred onto Luminex beads for use in a bead flow cytometric immunoassay. The novel one-step antibody coupling process increased assay sensitivity of a Luminex tumor necrosis factor immunoassay. These high-binding surfaces do not require prior incorporation of polyhistidine tags or posttreatments such as oxidation to achieve essentially irreversible binding of immunoglobulin G.