High-throughput optimization of surfaces for antibody immobilization using metal complexes.
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.
Muir
Ben W
BW
Bio-Layer Pty Ltd, Brisbane Technology Park, Unit 4, 26 Brandl st., Eight Mile Plains, 4113 QLD, Australia.
Barden
Michael C
MC
Collett
Simon P
SP
Gorse
Alain-Dominique
AD
Monteiro
Raisa
R
Yang
Liqun
L
McDougall
Nicole A
NA
Gould
Sharon
S
Maeji
N Joe
NJ
D000818:Animals; D000937:Antigen-Antibody Reactions; D001128:Argon / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods*; D002857:Chromium / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D008567:Membranes, Artificial; D019151:Peptide Library; D011137:Polystyrenes / Q000737:chemistry; D011145:Polyvinyls / Q000737:chemistry; D040081:Protein Array Analysis; D011506:Proteins / Q000276:immunology* / Q000378:metabolism; D012680:Sensitivity and Specificity; D005465:Fluorocarbon Polymers
D001128:Argon; D002857:Chromium; D007074:Immunoglobulin G; D008567:Membranes, Artificial; D019151:Peptide Library; D011137:Polystyrenes; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers