We report a novel high-throughput (HTP) protein chip platform, constructed on gold using self-assembly techniques, for conducting high quality antigen-antibody interactions. Biotinylated monolayers were used to immobilize a streptavidin surface with high packing density. This biocompatible platform was then used for detection of serum IgM antibodies. Serum samples of patients suspected to suffer from Lyme borreliosis were used to validate the protein chip platform using biotinylated peptide AAOspC8 molecules as the test probes. Various experimental parameters such as the effect of concentration of probes, targets, temperature of incubation, and their effect on the resulting signal-to-noise ratio are described in detail. Highly specific protein interaction data with a high signal-to-noise ratio were obtained with serum sample solutions as low as 1 microL/spot (1/10 diluted).

Abstract— The naphthalene‐sensitized formation of triplet excited chlorophyll‐a (Chl‐a) and all‐transß‐carotene has been studied by pulse radiolysis. The rate constants for transfer of triplet energy from naphthalene to Chl‐a and all‐transß‐carotene in benzene at 25°C are (3.6 ± 0.6)·109M‐1 s‐1 and (10.7 ± 1.2)·109M‐1 s‐1, respectively. The decays of the excited triplet states of naphthalene, Chl‐a and all‐transß‐carotene all follow a mixed first‐and second‐order mechanism. The first‐order rate constant for triplet decay is strongly dose dependent for naphthalene but only slightly dependent and independent of dose for Chl‐a and all‐transß‐carotene, respectively. The rate constants for triplet‐triplet annihilation are (1.4 ± 0.3)·109M‐1 s‐1 for Chl‐a and (3.6 ± 0.4)·109M‐1 s‐1 for all‐transß carotene. The nearly constant ratio k(ß‐carotene)/k(Chl‐a) for the bimolecular triplet energy transfer rate constants is discussed in terms of the molecular shapes of the two molecules. The energetics of the triplet‐triplet annihilation of all‐transß‐carotene are discussed, and it is proposed that production of the excited 1AB state may be a major route in the annihilation process.