Can Ozbal

Momentum Biotechnologies

Label-free in vitro potency assays are an emerging field in drug discovery to enable more physiological conditions, to improve the readout quality, and to save time. For this approach mass spectrometry (MS) is a powerful technology to directly follow physiological processes. The speed of this methodology, however, was for a long time not compatible...

Aim We describe the validation of an HPLC–MS/MS method to analyze ceftolozane and tazobactam simultaneously in saline matrixes. Materials & methods An Agilent 1260 HPLC interfaced to an Agilent 6470 triple-quadrupole mass spectrometer was used for quantification. A reverse-phase column running a gradient of water and acetonitrile containing 0.1% f...

Embodiments of the present disclosure present novel systems, devices and methods for an automated biological sample analysis using mass-spectrometry. The time from sample introduction to the reporting of data, in some embodiments, takes a relatively short amount of time (e.g., several minutes). In some embodiments, a biological sample to be analyze...

The invention features methods of making devices, or "platens", having a high-density array of through-holes, as well as methods of cleaning and refurbishing the surfaces of the platens. The invention further features methods of making high-density arrays of chemical, biochemical, and biological compounds, having many advantages over conventional,...

One embodiment of the invention provides a sample injection system including a vacuum source, a conduit in communication with the vacuum source, a fluid sensor configured to detect the presence of the fluid in the conduit, a sample loop in communication with the conduit; and a sipper in communication with the sample loop.

Aspects of the invention provide systems and methods for high-throughput screening of fluidic samples. In some embodiments, two chromatography columns are utilized in series. The first chromatography column can have a high affinity for phosphorylated compounds while the second chromatography column has a high affinity for one or more analytes of in...

Aspects of the invention provide systems and methods for high-throughput screening of fluidic samples. In some embodiments, two chromatography columns are utilized in series. The first chromatography column can have a high affinity for phosphorylated compounds while the second chromatography column has a high affinity for one or more analytes of in...

One embodiment of the invention provides a method of preparing an eluted sample containing salts or buffers from a liquid chromatography device for analysis by a mass spectrometry device. The method includes: continuously providing a non-polar solvent to the mass spectrometry device; receiving the eluted sample from the liquid chromatography device...

We describe here a high-throughput assay to support rapid evaluation of drug discovery compounds for possible drug-drug interaction (DDI). Each compound is evaluated for its DDI potential by incubating over a range of eight concentrations and against a panel of six cytochrome P450 (CYP) enzymes: 1A2, 2C8, 2C9, 2C19, 2D6, and 3A4. The method utilize...

The sirtuin enzymes, a class of NAD(+)-dependent histone deacetylases, are a focal point of epigenetic research because of their roles in regulating gene expression and cellular differentiation by deacetylating histones and a host of transcription factors, including p53. Here, the authors present two label-free screening methodologies to study sirt...

Histone acetyltransferases (HATs) catalyze the transfer of an acetyl group from an acetyl-coenzyme A donor molecule to specific lysine residues within proteins. The acetylation state of proteins, particularly histones, is known to modulate their intermolecular binding properties and control various cellular processes, most notably transcriptional a...

The evaluation of interactions between drug candidates and transporters such as P-glycoprotein (P-gp) has gained considerable interest in drug discovery and development. Inhibition of P-gp can be assessed by performing bi-directional permeability studies with in vitro P-gp-expressing cellular model systems such as Caco-2 (human colon carcinoma) cel...

A high-throughput online solid-phase extraction/tandem mass spectrometry (online SPE/MS/MS) system has been developed to support rapid evaluation of drug discovery compounds for possible drug-drug interaction (DDI). Each compound is evaluated for its DDI potential by incubating over a range of 8 test concentrations and against a panel of 6 cytochro...

A high-throughput mass spectrometry assay to measure the catalytic activity of UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase, LpxC, is described. This reaction is essential in the biosynthesis of lipopolysaccharide (LPS) of gram-negative bacteria and is an attractive target for the development of new antibacterial agents. The assay...

In this review various technologies and approaches for the utilization of mass spectrometry in high-throughput analyses are discussed. The use of quadrupole-based mass spectrometry in the screening of chemical libraries against enzymatic targets for the identification of inhibitors and/or activators is highlighted. The RapidFire mass spectrometry s...

Objective: Assessment of cytochrome P450 inhibition by NCEs has moved into earlier phases of drug discovery/development, and recent guidance from the FDA recommends routine in vitro IC50 determinations for at least six P450 isoforms. The resulting increase in the number of samples generated in P450 inhibition screens has led to a demand for higher...

Recently there has been a greater recognition of the importance of screening for time-dependent inhibition (TDI) of human cytochrome P450s earlier in drug discovery. This has lead to the demand for higher throughput analysis options for this assay. The objective of this study was to evaluate single vs multi-concentration methods for in vitro TDI sc...

The metabolic half-life or stability of a drug candidate has important pharmacokinetic and clinical significance, because it influences both oral bioavailability and plasma concentration of a compound, ultimately affecting efficacy. Large compound libraries and advancements in liquid handling have placed demands on the throughput, reliability and r...

An auto-injection system provides high throughput screening of fluidic samples. A sample injection valve has a first position which applies a reduced pressure to a sample sipper tube for aspirating a fluidic sample into the sample sipper tube, and a second position which delivers the fluidic sample to a sample supply loop. A column control valve ha...

Label-free mass spectrometric (MS) technologies are particularly useful for enzyme assay design for drug discovery screens. MS permits the selective detection of enzyme substrates or products in a wide range of biological matrices without need for derivatization, labeling, or capture technologies. As part of a cardiovascular drug discovery effort a...

Objective: Assessment of cytochrome P450 inhibition by NCEs has moved into earlier phases of drug discovery/development, and recent guidance from the FDA recommends routine in vitro IC50 determinations for at least six P450 isoforms. The resulting increase in the number of samples from P450 inhibition screens has lead to a demand for higher through...

Objective: Recently there has been a greater recognition of the importance of screening for time-dependent inhibition (TDI) of human cytochrome P450s earlier in drug discovery. This has lead to the demand for higher throughput analysis options for this assay. The objective of this study was to evaluate single vs multi-concentration methods for in v...

Several recent reports suggest that stearoyl-CoA desaturase 1 (SCD1), the rate-limiting enzyme in monounsaturated fatty acid synthesis, plays an important role in regulating lipid homeostasis and lipid oxidation in metabolically active tissues. As several manifestations of type 2 diabetes and related metabolic disorders are associated with alterati...

Small molecule high-throughput screening in drug discovery today is dominated by techniques which are dependent upon artificial labels or reporter systems. While effective, these approaches can be affected by certain experimental limitations, such as conformational restrictions imposed by the selected label or compound fluorescence/quenching. Label...

A high-throughput mass spectrometry assay to measure the catalytic activity of phosphatidylserine decarboxylase (PISD) is described. PISD converts phosphatidylserine to phosphatidylethanolamine during lipid synthesis. Traditional methods of measuring PISD activity are low throughput and unsuitable for the high-throughput screening of large compound...

Mass spectrometry is an emerging format for label-free high-throughput screening. The main limitation of mass spectrometry is throughput, due to the requirement to purify samples prior to ionization. Here the authors compare an automated high-throughput mass spectrometry (HTMS) system (RapidFire) with the scintillation proximity assay (SPA). The ca...

In accordance with an illustrative embodiment of the invention, a system for obtaining a fluidic sample from a container is presented. The container defines a volume for holding the sample, the volume being closed by a seal. An outer tube has a proximal end and a distal end, the proximal end shaped to permit piercing the seal. An inner tube has an...

Cytochrome P450 3A4 (CYP3A4) is often regarded as the most important P450 responsible for drug metabolism. In addition to CYP3A4, CYP3A5 represents at least 50% of the total hepatic CYP3A content in those polymorphically expressing CYP3A5, and may be the most important genetic contributor to interindividual and interracial differences in CYP3A depe...

A system and method high throughput sample preparation and analysis using column chromatography. Each port in a plurality of ports has a port input that interfaces with a first fluid source and a port output. A fluidic circuit is coupled to each port output and to a second fluid source, the fluidic circuit for controlling fluid flow from the plural...

A system and method high throughput sample preparation and analysis using column chromatography. Each port in a plurality of ports has a port input that interfaces with a first fluid source and a port output. A fluidic circuit is coupled to each port output and to a second fluid source, the fluidic circuit for controlling fluid flow from the plural...

Mass spectrometry-based screening can be applied to a wide range of targets, including those intractable targets that use substrates such as lipids, fatty acids, phospholipids, steroids, prostaglandins, and other compounds not generally amenable to conventional screening techniques. The major limitation to this approach is throughput, making HTS vi...

The invention features methods of making devices, or “platens”, having a high-density array of through-holes, as well as methods of cleaning and refurbishing the surfaces of the platens. The invention further features methods of making high-density arrays of chemical, biochemical, and biological compounds, having many advantages over conventional,...

In the present report, we describe a fluorescence-based method capable of measuring benzo[alpha]pyrene diolepoxide (BPDE) adducts in intact genomic DNA, with a sensitivity of a few hundreds copies per cell. The assay is based on cryogenic laser-induced fluorescence technology at liquid nitrogen temperatures, coupled with an intensified charge-coupl...

The ubiquitous environmental carcinogen benzo[a]pyrene (BaP) is metabolized in vivo in humans to its ultimate carcinogenic form of 7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE). Mouse skin tumorigenicity studies indicate that the (7R,8S,9S,10R) enantiomer of BPDE, (7R,8S)-dihydroxy-(9S,10R)-epoxy-7,8,9,10-tetrahydrobenzo[a]pyren...

Tributyltin (TBT) and dibutyltin (DBT) were analyzed in sediment samples collected from intertidal locations in Portland and Boothbay Harbor, Maine (USA) in 1990 and 1992. Surface sediment TBT concentrations ranged from 24 to 12 400 ng gm(-1) (dry wt basis). Sediments with the highest TBT concentrations were associated with shipyard hull washing/re...

The effect of cell replication on histone-carcinogen adducts was investigated by determining the specific adduct levels as a function of time following carcinogen treatment of human TK6 cells grown in culture. Core histones isolated from cells treated with aflatoxin B1 or r-7,t-8 dihydroxy-t-9,t-10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene exhibited...

The throughput of in vitro ADME analysis is gaining an almost equal footing with data quality as a determinant of laboratory workflow. These assays have traditionally utilized tandem MS, which is limited by its requisite MRM method development that requires several minutes per sample of processing time. The use of accurate mass offered by high reso...

MS based cytochrome p450 assays are becoming commonplace in early drug discovery efforts. Growing throughput demands coupled with limited supply in single donor material has necessitated the use of pooled liver microsomes in standard protocols. The use of mixed donor enzymes cannot account for the polymorphisms in multiple p450 enzymes across the p...

The metabolic half-life or stability of a drug candidate has important pharmacokinetic and clinical significance, because it influences both oral bioavailability and plasma concentration of a compound, ultimately affecting efficacy. Large compound libraries and advancements in liquid handling have placed demands on the throughput of in vitro metabo...