How Electronic Laboratory Notebooks Aid in the Development of Bioanalytical Assays for Large Molecule Detection by LC/MS

The use of electronic laboratory notebooks (ELNs) is becoming more popular among research and development (R&D) organizations. These electronic platforms optimize workflow and expedite data analysis while offering intellectual property (IP) protection, audit trail recording, and business rule enforcement. With the evolution of science and technology over the past 20 years, many lab operations are well developed and understood, but data processing, review, and reporting can take up to triple the time it takes for the operation itself.1 Bioanalytical organizations are keen to reap the benefits by capitalizing on ELNs for use in these already developed research areas.

Harnessing data

With strictly regulated processes and intensive quality audits, there are several ways an ELN can help bioanalytical laboratories increase efficiency; in particular, bioanalytical operations are looking to improve productivity and reduce overall costs. A good platform improves ways of capturing and organizing structured and unstructured data. It enhances workflow and process management, automatically enforces quality assurance (QA), and offers superior reporting capabilities. These benefits can be realized by mature platforms and the development of new ones.

Bioanalytical laboratories are eagerly investigating methods to better quantify them, as large molecules gain importance as therapeutic agents. Until recently, ligand binding assays (LBAs) have been the most common approach to large molecule quantitation for bioanalysis. Developments in LC/MS instrumentation and technique have now made LBAs a viable alternative for quantitation of large molecule proteins.2 While analytical technologies are being greatly improved, the usual regulated data collection and review process takes exceptionally longer in comparison to the technology itself.1 This drastically slows down the innovation process and increases costs. ELNs reduce the time it takes for data collection and review, and also enforce limits for data entry, automated data calculation, and analysis and auditing.

Meeting LC/MS challenges

Rolling out new LC/MS technology for large molecule quantitation does have its challenges, including increased sensitivity and selectivity of the assay, optimization of sample preparation, and obtaining reproducible and accurate results. Sample preparation is greatly important for increasing the sensitivity of an assay and is often the rate-limiting step in bioanalysis.3 Preparation technique depends largely on the molecular weight of the protein. The effect of sample containers must also be considered. Studies have shown a loss of sensitivity due to the propensity of a protein or peptide anlayte to nonspecifically bind to the container surface.4 Optimizing the preparation of the sample will minimize this variability and aid in increasing sensitivity of the assay.

An ELN may be used to monitor the type of sample preparation based on any business rule that is set during assay development. It will control preparation while monitoring that the selections made pass business rules, flagging any exceptions simultaneously; for example, the molecular weight of a protein being analyzed can determine the necessary or advised preparation technique(s). Preparation steps can be flagged when inappropriate for the molecular weight range of the specified protein. Additionally, the container for sample preparation, preparation buffers, and reagents can also be chosen based on defined business rules. Tailored drop-down lists can be dictated based on the properties of the protein, such as molecular weight, protein type, etc.

Generating quality data

Reproducibility of data is paramount in order for this new method to be an acceptable technique for large molecule quantitation. Moreover, accurate data collection and analysis are the most time-consuming part of method development. An ELN offers numerous advantages to expedite data capture and improve data quality and compliance. Simultaneously it provides quality control (QC) in real time. Transcription errors are very common in a paper-based world. An ELN approach means data can be read digitally, directly off of an instrument—this eliminates the need for transcription at all.

By applying business rules to sample data, an ELN enforces the rules and prioritizes values that fall outside of approved limits and acceptance criteria. Traditional QC is performed after data have been acquired, with errors identified and addressed after they occur. The use of an electronic system incorporates QC checks into the workflow from beginning to end. When data fall outside regulated success criteria, workflow controls that are already in place can dictate when a test passes or fails. The ELN also highlights if an instrument needs to be calibrated, when reagents are expired, if a test needs to be repeated or restarted, or any other business requirements.

Validating methods once developed is a time-consuming and difficult step due to the postanalysis run review. An ELN provides validation templates, equivalent to a paper form, which may be subjected to a specific development and validation procedure. These primary templates can then be used to create method-specific secondary templates, which are created quickly from a trustworthy source.1 Many values in a secondary template are not changes to the primary template, but simply specific to the method being tested, and thereby do not require further review.1 Of course, these templates have the same QC checks built into them as described above, ensuring every step of the process is being evaluated and verified in real time.

Embracing technological advantage

The bioanalytical laboratory is responsible for the assessment of drug safety and bioequivalence.1 New Microsoft-based technologies offer enormous possibilities in the bioanalytical world. Before we can use these methods as an industry standard, they must be developed, tested, and validated. The process of developing these methods has been slow and cumbersome due to regulatory pressures. An ELN can better equip these laboratories with more efficient data collection, data analysis, and quality data checks in real time, to drastically improve the whole process.

References

  1. Shoup, R.E., Beato, B.D. et al. Implementation of an electronic laboratory notebook to accelerate data review in bioanalysis. Bioanalysis  2013, 5(13), 1677‒89.
  2. Knutsson, M., Schmidt, R. et al. LC-MS/MS of large molecules in a regulated bioanalytical environment—which acceptance criteria to apply? Bioanalysis  2013, 5(18), 2211‒14.
  3. Souverain, S., Rudaz S. et al. Restricted access materials and large particle supports for online sample preparation: an attractive approach for biological fluids analysis. J. Chromatogr. B  2004, 801(2), 141‒56.
  4. Ramagiri, S., Garofolo, F. Large molecule bioanalysis using Q-TOF without predigestion and its data processing challenges. Bioanalysis  2012, 4(5), 529‒40.

Laura Swift is Senior Solutions Consultant, IDBS, 2 Occam Ct., Surrey Research Park, Guildford, Surrey GU2 7QB, U.K.; tel.: +44 1483 595 000; www.idbs.com/

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