Non-GLP Bioanalytical Methods

In discovery bioanalysis, the lab is often blind to analyte structure. To proceed swiftly and intelligently to predict drug uptake and elimination in support of compound identification and screening, we utilize a research grade assay approach via LC-MS/MS to quickly analyze samples for compound ranking. Once a lead candidate is selected, these methods can be transferred to method development for GLP bioanalytical testing of nonclinical and clinical samples in support of your regulatory submissions.

Our Discovery Bioanalysis Approach

Discovery bioanalysis is not “quick and dirty”; on the contrary, it requires a carefully chosen, well-planned approach to analyze samples without a validation. To start, a default protein precipitation LC-MS/MS method is applicable to most small molecule compounds, eliminating the need for method development or qualification. Features of the protein precipitation methodology include:

  • Assay range of 1–5000 ng/mL (default; can be modified as required)
  • Front and back calibration standards consisting of nine different concentrations
  • Sample aliquot volume default of 10 µL of biofluid (particularly important for serial mouse sampling)
  • Addition of gradient selection based on retention time

The use of bracketing calibration standards is crucial in discovery bioanalysis to control potential divergence (drift) that can occur due to buildup of matrix components or dosing vehicle. This effort is further aided by using small sample volumes, which minimize these adverse effects, plus our use of the latest technologies, such as UPLC and Tandem MS (LC-MS/MS), to achieve appropriate sensitivity even with such small sample aliquots.

Research-grade Assays

The key to balancing the competing demands of data quality, speed of execution, and cost in discovery bioanalysis is a practical understanding of the data quality required to satisfy experimental objectives. Charles River offers a suite of standardized, fit-for-purpose non-GLP research-grade assays (RGA) to address client needs in the discovery bioanalysis space across the drug continuum. Additionally, for studies with multiple matrices, we can flexibly work on alternative assay scenarios to effectively manage cost and minimize time while ultimately satisfying data quality requirements. We can perform studies using client-defined procedures and acceptance criteria to ensure data comparability, consistent with our philosophy of providing services as a true extension of your internal capabilities and processes.

Diagnostic Probes

Experience has taught us that great design must be followed by great execution when performing discovery bioanalysis. To ensure this, we have designed numerous diagnostic probes embedded into our sample analysis procedures to assess and demonstrate the production of high-quality data. Diagnostic probes are further used to identify, diagnose, and troubleshoot areas where discovery bioanalytical data quality may be compromised, enabling us to solve problems and re-establish quality quickly. Examples of these probes include:

  • Multiple internal standards (to accommodate a range of retention times and to assess suppression)
  • Multiple transitions for each analyte
  • Matrix effect probe (to determine if matrix or vehicle may be causing quantitation interferences)
  • Discovery bioanalysis stability probe (to assess short-term stability of the analyte in study matrix)

The biopharmaceutical industry relies heavily on drug discovery to reduce time and cost in finding a successful drug candidate; therefore, we continually improve to drive quality and productivity so that discovery bioanalysis can generate data with tighter acceptance criteria and deliver higher quality data even faster. The data and knowledge generated can provide feedback on key in vitro assays used to define the medicinal chemistry space and through in vivo studies provide an early understanding of the relationship between concentration and efficacy. Learn more about how bioanalysis plays a pivotal role in supporting pharmaceutical research from drug discovery and development through to market and how chemists are engaged every step of the way.

Frequently Asked Questions (FAQs) for Discovery Bioanalysis

  • What is discovery bioanalysis?

    Discovery bioanalysis provides rapid non-GLP bioanalytical data, allowing an evaluation of the PK characteristics of potential drug candidate compounds while minimizing time and cost during the early and late stages of drug discovery.

  • What is a Research Grade Assay (RGA)?

    The scientific approach to provide bioanalytical data with minimum method development necessary to yield data with the quality attributes necessary to answer the questions being asked in the lead compound identification and optimization phase prior to regulated nonclinical studies begin.

  • What data should I expect from a discovery bioanalysis pharmacokinetics project?

    Standard data generated from a research grade assay includes non-compartmental data, time to Cmax (tmax), Cmax, t1/2 elimination (terminal elimination half-life) and AUC (area under the curve).