Automation of the anthrone assay for carbohydrate concentration determinations.

Reported is the adaptation of a manual polysaccharide assay applicable for glycoconjugate vaccines such as Prevenar to an automated liquid handling system (LHS) for improved performance. The anthrone assay is used for carbohydrate concentration determinations and was scaled to the microtiter plate format with appropriate mixing, dispensing, and measuring operations. Adaptation and development of the LHS platform was performed with both dextran polysaccharides of various sizes and pneumococcal serotype 6A polysaccharide (PnPs 6A).

An integrity assay for a meningococcal type B conjugate vaccine.

The development of an analytical procedure for the evaluation of a conjugate vaccine's structural wholeness or integrity is described. The principle component of the vaccine was the N-propionylated group B meningococcal polysaccharide (NPr-GBMP) covalently attached to a carrier protein. The goal of the procedure was to determine whether any whole polysaccharide, oligosaccharide, or monosaccharide, from minute to moderate levels, became detached off the conjugate.

Complete structure elucidation of a globular protein by particle beam liquid chromatography-Fourier transform infrared spectrometry and electrospray liquid chromatography-mass spectrometry. Sequence and conformation of beta-lactoglobulin.

The advantages to the use of both mass spectrometry (MS) and Fourier transform infrared spectrometry (FT-IR) in combination for the structural characterization of the tryptic digest of a model globular protein is demonstrated. HPLC has been interfaced to both spectroscopic techniques and has provided a high degree of structural detail for the target protein. beta-Lactoglobulins A and B were digested with trypsin and chromatographed with narrow-bore, reversed-phase HPLC.

Sulfate detection in glycoprotein-derived oligosaccharides by artificial neural network analysis of Fourier-transform infrared spectra.

We report the use of an artificial neural network to analyze the fingerprint region of Fourier-transform infrared (ir) spectra of oligosaccharides for the presence of sulfate groups. This assay can rapidly and nondestructively detect the presence of sulfate in as little as 1 nmol (approximately 2 micrograms) of a glycoprotein-derived monosulfated decasaccharide. The neural network was trained to recognize the presence of sulfate groups by presenting it with 45 ir spectra of sulfated and nonsulfated mono- and oligosaccharides.