Biomedical subjects

P G Mattingly

Publications and source records attributed to P G Mattingly.

23 records · Page 2Linked to original sources

PubMedDate not supplied

A convenient method for the preparation of hapten phosphoramidites.

Automated incorporation of haptens at the 3' or 5' end of oligonucleotides required the preparation of the corresponding hapten phosphoramidites. The requisite 1,3-diol framework was prepared in two steps from a carboxylic acid precursor first by a two-carbon homologation using Meldrum's acid to form the corresponding 3-oxo ester and then subsequent reduction to the diol. The primary alcohol was protected with a dimethoxytrityl group, while the secondary alcohol was converted to the reactive phosphoramidite.

Automation↗

PubMedDate not supplied

Characterization of protein-hapten conjugates. 2. Electrospray mass spectrometry of bovine serum albumin-hapten conjugates.

Fifteen hapten-bovine serum albumin (BSA) conjugates were prepared from five commercially available activated haptens. Each hapten was coupled to BSA at three different ratios. The conjugates were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two mass spectrometry (MS) methods: matrix-assisted laser desorption ionization (MALDI) and liquid chromatography-electrospray ionization (LC-ESI). SDS-PAGE was useful in detecting protein cross-linking, but not assessing hapten density. MALDI-MS and LC-ESI-MS gave comparable qualitative results, but LC-ESI-MS provided a clearer representation of the distribution of hapten-protein species present in the conjugates. Conjugate species substitute with up to 25 haptens per BSA were recorded by LC-ESI-MS.

Animals↗

PubMedDate not supplied

Immunoassay reagents for thyroid testing. 2. Binding properties and energetic parameters of a T4 monoclonal antibody and its Fab fragment with a library of thyroxine analog biosensors using surface plasmon resonance.

A library of 16 thyroxine analog biosensors were prepared for use on the BIAcore surface plasmon resonance instrument. An anti-thyroxine monoclonal antibody and its Fab fragment were tested with each biosensor to generate the association and dissociation rate constants, equilibrium association constants, and Gibbs free energy values for characterizing the binding event.

Animals↗

PubMedDate not supplied

Immunoassay reagents for thyroid testing. 3. Determination of the solution binding affinities of a T4 monoclonal antibody Fab fragment for a library of thyroxine analogs using surface plasmon resonance.

A library of thyroxine analogs and tracers was prepared, and their solution binding affinities for an anti-T4 Fab fragment were determined using a single high-density L-T4 biosensor surface in a BIAcore surface plasmon resonance instrument. The high-density L-T4 analog biosensor was calibrated by determination of the initial binding rate was of known concentrations of free anti-T4 Fab fragment in solution to the biosensor surface. A range of individual thyroxine analog and tracer concentrations was subsequently mixed with a fixed concentration of anti-T4 Fab fragment. The concentration of free anti-T4 Fab fragment in each solution at equilibrium was determined, and the equilibrium dissociation constant (KD) for each case was derived. The KD values determined in solution are compared to values determined by a direct kinetic analysis on the BIAcore instrument using individual biosensor surfaces.

Acridines↗

PubMedDate not supplied

Surface plasmon resonance (SPR) as a tool for antibody conjugate analysis.

Surface plasmon resonance (SPR) analysis was used to assess the immunoreactivity of anti-biotin (4) and anti-fluorescein (5) monoclonal antibody after conjugation with the N-hydroxysuccinimide ester of acridinium-9-carboxamide 1. Only minor changes in the apparent equilibrium dissociation constants of the antibody conjugates for their ligands resulted from the conjugation process. However, comparison of the initial binding rate of the conjugates with their ligands with those of the unmodified antibodies over a range of concentrations showed that the antibody conjugates were partially inactivated. The anti-fluorescein conjugates retained at least 90% of their immunoreactivity over the range of modification tested, while anti-biotin conjugates showed a progressive loss of reactivity with increased substitution by the label.

Acridines↗