In this study, the authors report that “The equivalence of RF-MS/MS to traditional LC/MS/MS was evaluated with 280 proprietary compounds, which were assayed to determine the inhibition of CYP-2D6, -2C9, or -3A4. The samples were split into two separate 384-well plates and analyzed on either the RF-MS/MS or LC/MS/MS platforms. The IC50 concentrations for the proprietary compounds were calculated and plotted in Excel to determine the correlation of the data between the two platforms. clogP values for compounds were calculated using BioByte version 4.71 (BioByte, Claremont, CA, USA) and pKa values were calculated using Molsoft ChemistPro 3.6-1g (Molsoft, La Jolla, CA, USA)”.

Here’s the link to the article:
[Citation: Brown, A., Bickford, S., Hatsis, P., Amin, J., Bell, L. and Harriman, S. (2010), High-throughput analysis of in vitro cytochrome p450 inhibition samples using mass spectrometry coupled with an integrated liquid chromatography/autosampler system. Rapid Communications in Mass Spectrometry, 24: 1207–1210. doi: 10.1002/rcm.4461]

The Situation: A small pharmaceutical company did not have the in-house MS analysis capacity to routinely analyze P450 inhibition screens. They wanted a cost effective out-sourcing solution that integrated seamlessly with their current workflow and also provided rapid data turnaround to assure steady progress in their drug discovery programs. Seamless integration for this client meant that they retained full control of their assay protocols and compound identities as well as flexibility in assay plate format.

Addressing the Challenges: Could the RapidFire service be used to analyze plates from their current assays without any further modifications to their protocols? The RapidFire team worked with the client to provide an evaluation of the RapidFire CYP inhibition service that would assure them that this service could provide consistent data rapidly and integrate seamlessly into their current workflow.

Results: The client continues to utilize the BIOCIUS RapidFire CYP Inhibition Service on a routine basis by sending quenched assay samples to BIOCIUS for analysis. Data from P450 inhibition assays is sent back to the client within 3 days of receipt and often faster. By utilizing this analysis service the client was able to get the data they need while fully controlling assay protocols and preparation, compound identity, and plate formats without delays in their drug discovery programs.

The Situation: A large pharmaceutical company was consolidating global P450 inhibition screening operations to a single location. This single facility now needed to meet the requirements of quickly and efficiently performing these assays at a capacity several fold higher than their current abilities without significantly altering their current assay protocols including the acquiring of data from full IC₅₀ curves for several CYP isoforms. The global capacity analysis needs for this assay was estimated to be in excess of 500,000 samples per year.

Addressing the Challenges: The RapidFire team worked with the client to customize the existing RapidFire CYP inhibition analysis methods to meet their requirements. The new methods not only needed to be quick and efficient, but they also needed to measure up to an extensive validation of cross platform equivalence (in-house data vs. RapidFire) evaluated by analyzing a data set containing tens of thousands of wells over a time period of several months.

Results: The RapidFire platform and newly developed methods successfully met the client’s high expectations by increasing analysis speed and sample volume capacity without sacrificing data integrity. Initially the client utilized the BIOCIUS RapidFire CYP Inhibition Service by sending quenched assay samples in high volume to BIOCIUS for analysis. After a period of time the client worked closely with the RapidFire team to install a RapidFire MS system in their facility and transition the sample analysis in-house.

For more information please visit www.BIOCIUS.com

Check out this interesting article about the challenges facing researchers in the field of pharmaceutical analysis; visit the BIOCIUS website to see how RapidFire can efficiently address these challenges.

On June 23rd BIOCIUS held an open house to celebrate the move to the new facility!

It was a beautiful evening in Wakefield, and attendees enjoyed good company, food and the weather on BIOCIUS’ patio!

On June 24th, BIOCIUS employees: Lauren Frick, Margaret Chute, and Lauren Frick (picture on left) represented BIOCIUS in the 27th annual J.P. Morgan Corporate Challenge on the Boston Common.  The challenge is a 3.5 mile loop and boasted 708 companies from the Boston area.  Lauren Frick of BIOCIUS said of the event, “We were glad the weather held off; the corporate challenge was a fun event.  There was an impressive turnout for a great cause, and we were very proud to represent BIOCIUS.”

In addition to the booth and hospitality suite, RapidFire was included in an oral presentation by Panos Hatsis, “Reducing Bottlenecks in ADME Sample Analysis using Solid Phase Extraction with a Quadrupole Time-of-Flight Mass Spectrometer” and  multiple posters: “Evaluation of Accurate Mass TOF-MS in High Throughput PAMPA Screening”, “Comparison of LC/MS/MS and a Fully Integrated Autosampler/Solid Phase Extraction System for the Analysis of Permeability Samples”, and “Development of a High-Throughput MS-Based Bioanalytical Method for Assessment of P-Glycoprotein Inhibition”

For any questions about BIOCIUS or presentations/posters at ASMS please visit our website www.BIOCIUS.com or contact us at: 781-928-2700, info@BIOCIUS.com

See you next year in Colorado!

Hospitality suite 251A for:

• Hors d’oeuvres
• Refreshments
• Foosball
• Prizes!

Also check out RapidFire presentations:

Monday

Oral Presentation 8:50 a.m., Hall 2

Reducing Bottlenecks in ADME Sample Analysis using Solid Phase Extraction with a Quadrupole Time-of-Flight Mass Spectrometer
Panos Hatsis ¹; Michelle Romm²; Vaughn Miller²; Jakal Amin¹; William A. Lamarr²; Can “Jon” Ozbal²; Shawn Harriman¹¹Novartis Institutes for Biomedical Research, Cambridge , MA; ²BIOCIUS Lifesciences, Woburn, MA

Novel Aspect: SPE-QqTof in an integrated ADME workflow with specificity and efficiency in a wide range of ADME assays.

Tuesday

Poster Session TP21, #498 & #500

Evaluation of Accurate Mass TOF-MS for Use in High Throughput PAMPA Screening
Michelle Romm ; Nikunj Parikh; Vaughn Miller; William A. Lamarr; Can “Jon” Ozbal BIOCIUS Life Sciences, Inc., Woburn, MA
Novel Aspect: Use of an integrated SPE/Q-ToF system with generic methods for improving accuracy and efficiency in PAMPA sample analysis.

Time Slot/Poster Number: 500 Session: Drug Metabolism: High Throughput
Comparison of LC/MS/MS and a Fully Integrated Autosampler/Solid Phase Extraction System for the Analysis of Permeability Samples
Adam C Amaral; Christopher Caldwell; Panos Hatsis ; Jakal Amin; Shawn Harriman Novartis, Cambridge, MA
Novel Aspect: The successful development and implementation of a complete high-throughput workflow process for the routine analysis of permeability samples.

Thursday

Poster Session ThP24, #572

Session: High Throughput Analysis/Robotics
Development of a High-Throughput MS-Based Bioanalytical Method for Assessment of P-Glycoprotein Inhibition
Andrew Wagner^{1, 2}; Janet Kolb^{1, 2}; John Herbst^{1, 2}; Tatyana Zvyaga^{1, 2}; Conway Charlie^{1, 2}; Harold Weller^{1, 2}; Wilson Shou ^{1, 2}; Can “Jon” Ozbal^{3 1}Bristol-Myers Squibb Company, Wallingford, CT; ²Bristol-Myers Squibb Company, Princeton, NJ; ³BIOCIUS Life Sciences, inc, Woburn, MA
Novel Aspect: By facilitating ultra-fast bioanalysis, the RapidFire™-MS/MS enables identification of potential drug-transporter interactions in the early stages of compound liability screening.

Hope to see you in Salt Lake City!

http://biocius.com/EpigeneticsWebinar.html

Are you working on sirtuins, HDACs, methyltransferases or other difficult screening programs?

As an alternative to fluorescence-tagged peptide assays in epigenetic research, BIOCIUS would like to share a unique approach to this challenging target class. In this complimentary 60 minute webinar, BIOCIUS VP Dr. LaMarr will present data from label-free screening projects for sirtuin histone deacetylases using a universal method developed by BIOCIUS scientists.

Register now for this webinar.  Topics for discussion include:

• a universal label-free screening method to assay the sirtuin class
  of NAD+ dependent histone deacetylases.

• the applicability of this assay to multiple members of the sirtuin
  family as well as multiple peptide and protein substrates.

• comparisons between classical peptide based assays and the direct measurement of the unique O-acetyl-ADP-ribose product.

In a recent report (reference below), Dr. Jonathan Baell and Dr. Georgina Holloway describe a series of “filters” that can be used to weed out compounds that have historically presented themselves as false positives in a multitude of HTS assays, to help researchers prevent the wasteful expenditure of resources on compounds that may be worthless.  They call these promiscuous positives PAINS (pan-assay interference compounds) and the filters that identify such PAINS are actually text files that can be incorporated into software used to sift through chemical libraries.

To come up with the different PAINS classes, Baell and Holloway analyzed data from previous chemical screens and painstakingly identified specific compound substructures that were associated with more positives than statistically likely.  For example, cyano-imines were identified as a class of PAINS because their susceptibility to reaction with the nucleophillic groups on proteins has previously made them appear to have specific inhibitory properties that were in fact artifactual.  Rhodanines also stuck out as potential PAINS because their color, metal chelating properties, and high reactivity with proteins has convoluted a large number assay results.

In total, over 400 classes of PAINS were identified, and the complete list of filters was made publicly available on February 4^th 2010 through the online publication in the Journal of Medicinal Chemistry.  Baell said he was contacted by a number of pharmaceutical companies within 48 hours of publishing the filters.  So, it appears as though the potential for these filters to help assess chemical libraries and/or hits is gaining recognition.  Hopefully, these filters prove to be useful, facilitate the identification of compounds worthy of development, and have a positive impact on numerous HTS efforts.

The article can be accessed at J. Med. Chem., 2010, 53 (7), 2719-2740, and I recommend it to all researchers interested in screening.

Peter T. Rye

http://pubs.acs.org/doi/abs/10.1021/jm901137j

When: April 11-15, 2010

Visit BIOCIUS at booth # 913 and also check out RapidFire presentations at SBS:

RapidFire Tutorial presented by Dr. Vaughn P. Miller: “Increasing the Speed and Efficiency of ADME Screening Assays by Utilizing RapidFire® MS Technology” Wednesday,April 14, from 8:00 a.m. – 8:45 a.m. Room 121AB Refreshments served

RapidFire Presentation by Dr. William LaMarr: “A Label-Free Screening Approach for Sirtuin Histone Deacetylases.” Thursday, April 15, from 10:30 a.m. – 11:00 a.m.

RapidFire Poster: A100 “A High-Throughput In Vitro ADME Assay Panel: Incorporation of SPE/MS/MS Into Standard Laboratory Workflow”

We hope to see you there!