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Biomedical subjects

D Dogruel

Publications and source records attributed to D Dogruel.

5 recordsLinked to original sources
PubMed1997

BIA/MS: interfacing biomolecular interaction analysis with mass spectrometry.

Biomolecular interaction analysis (BIA) which utilizes surface plasmon resonance (SPR) detection of affinity-captured analytes has been interfaced with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI). Femtomole quantities of a peptide, myotoxin a, were detected by direct MALDI analysis of sensor chips used during BIA of a polyclonal anti-myotoxin a IgG/myotoxin a system. Further, different interactive surfaces (flow cells) present on a single biosensor were targeted individually for mass spectrometric analysis. System compatibility of the combined approach was demonstrated with sensitivities, detection limits, and analytical performances comparable to those intrinsic to the individual analyses. The combined approach unites the real-time capabilities of SPR-based BIA with the qualitative specificity of mass spectrometry.

Biosensing Techniques↗
PubMed1995

Rapid tryptic mapping using enzymatically active mass spectrometer probe tips.

A method has been developed for rapid, sensitive, and accurate tryptic mapping of polypeptides using matrix-assisted laser desorption/ionization time-of-flight mass analysis. The technique utilizes mass spectrometer probe tips which have been activated through the covalent immobilization of trypsin. The enzymatically active probe tips were used for the tryptic mapping of chicken egg lysozyme and the results compared with those obtained using either free trypsin or agarose-immobilized trypsin. A significant increase in the overall sensitivity of the process was observed using the active probe tips, as well as the production of more characteristic proteolytic fragments and the elimination of background signals due to the autolysis of the trypsin. Further, probe tip digestions were found to be rapid and convenient.

Amino Acid Sequence↗
PubMed1995

Peptide characterization using bioreactive mass spectrometer probe tips.

A method has been developed for the rapid and sensitive mass spectrometric characterization of peptides. The approach uses bioreactive mass spectrometer probe tips, incorporating covalently bound enzymes, which are capable of modifying biomolecules for analytical purposes. In the demonstrated cases, enzymatic proteolysis is initiated upon application of analyte to the probe tips, time is allowed for digestion, and the products are analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The probe tips have been used for proteolytic mapping and partial sequence determination of picomole quantities of peptide. Analysis times were approximately 30 min. Two methods of database search were utilized. The first used limited peptide sequence information and parent molecular weight, while the second used exclusively the molecular weights of a number of endoproteolytic fragments. A simple method of comparing the match of experimental data for a tryptic digest with the results of a search is described.

Amino Acid Sequence↗
PubMed1994

Mass determination of human immunoglobulin IgM using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Monoclonal human immunoglobulin IgM has been analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Using a matrix of sinapinic acid with a laser wavelength of 355 nm, positive ions with 2 to 6 charges were observed in the mass spectra. Two major molecular species were present. The molecular weights estimated from the multiply-charged signals were found to decrease systematically with decrease in the charge state of the ion. This effect was determined to stem from the non-linear influence of the initial kinetic energy of desorption on the final kinetic energy of the multiply-charged species. A method of successive approximations, with the molecular weights and desorption velocities of the analytes as variables, was used to correct for these non-linearities. Mean molecular weights for the two IgM species were found to be 939,000 +/- 2000 and 982,000 +/- 2000 Da. An initial desorption velocity of 565 +/- 10 ms/s was determined for both.

Humans↗