High-throughput optimization of surfaces for antibody immobilization using metal complexes

Muir B.W., Barden M.C., Collett S.P., Gorse A.-D., Monteiro R., Yang L., McDougall N.A., Gould S. and Maeji N.J. (2007) High-throughput optimization of surfaces for antibody immobilization using metal complexes. Analytical Biochemistry, 363 1: 97-107. doi:10.1016/j.ab.2007.01.015


Author Muir B.W.
Barden M.C.
Collett S.P.
Gorse A.-D.
Monteiro R.
Yang L.
McDougall N.A.
Gould S.
Maeji N.J.
Title High-throughput optimization of surfaces for antibody immobilization using metal complexes
Journal name Analytical Biochemistry   Check publisher's open access policy
ISSN 0003-2697
Publication date 2007-04-01
Sub-type Article (original research)
DOI 10.1016/j.ab.2007.01.015
Volume 363
Issue 1
Start page 97
End page 107
Total pages 11
Language eng
Subject 1303 Specialist Studies in Education
1304 Biophysics
1312 Molecular Biology
Abstract 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.
Keyword Antibody immobilization
ELISA
High throughput
Metal complexes
Surface discovery
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
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Citation counts: TR Web of Science Citation Count  Cited 16 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 19 times in Scopus Article | Citations
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