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Biomedical subjects

James F LeBlanc

Publications and source records attributed to James F LeBlanc.

2 recordsLinked to original sources

ProteinChip array-based amyloid beta assays.

Amyloid-beta (A beta) fragments are found in plaques of patients with Alzheimers. Three secretases cleave the amyloid precursor protein, producing multiple A beta fragments that accumulate in the brain and fluids of patients with Alzheimers. A beta peptides are difficult to detect using standard methods because of their small size and multiple isoforms. However, multiple peptide fragments can be detected using a single ProteinChip Array-Based assay. Specific antibodies recognizing various amyloid epitopes are immobilized on a ProteinChip Array. Crude samples, such as tissue lysates, serum, cerebral spinal fluid (CSF), or cell culture media, are applied to the antibody-coated arrays. A beta peptides are specifically retained by the antibody, whereas other sample components are removed by washing. The multiple peptide fragments are detected by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS), which can easily resolve the different fragments because of the corresponding changes in peptide mass.

Alzheimer Disease↗

DNA affinity capture and protein profiling by SELDI-TOF mass spectrometry: effect of DNA methylation.

We report here that surface enhanced laser desorption/ionization-time of flight (SELDI-TOF) mass spectrometry, as performed on a Ciphergen Biosystems ProteinChip System, can be used in conjunction with DNA affinity capture (DACA) to study specific DNA-protein binding. Using DNA molecules bound to a surface, sequence-specific interactions can be detected as demonstrated by a mutation affecting the binding profile for TBP, a transcription factor. Also, a comparison between methylated and unmethylated promoter-containing DNA fragments shows numerous binding profile differences over a mass range extending to >60 kDa. The binding of several proteins is inhibited by methylation of the DNA.

Base Sequence↗