Assay Development

The art in Multiplexed Assays

Here at Protavio we know how to build assays for any type of applications including biomarker discovery, pathway analysis, quality control, PK/PD, screening studies etc. Using diverse coupling techniques, we can immobilise many different types of molecules (antibodies, affimers, proteins, etc) and employ detection schemes (sandwich assay, single antibody, direct labeling etc) that fit your project specification. If large scale production is needed, three different types of robots can be programmed to meet your production requirements. Following EMA, FDA, and CLSI’s guidelines we assess assay performance for Limits of Detection, precision, accuracy, linearity, and recovery.

Our process explained:

Step1: Select Analytes

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You can select any analyte even if antibodies are not available. Alternatively use our Megaplex™-1000 or focused panels to generate new hits, or consult our bioinformaticians for network-based biomarker analysis. We have more than 10 years experience on biomarker measurements, so feel free to contact us to discuss the selection process

Step2: Antibody Selection or Production

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Our team will select antibodies from (i) our validated antibody library, (ii) our trusted antibody suppliers, or (iii) we will produce polyclonal/monoclonal antibodies and standards for a de-novo assay development.

Step3: Antibody Evaluation

Smart Antibody Evaluation against whole proteome.

All antibodies are rigorously evaluated with our CROSS-PLEX platform where an antibody is checked against the whole proteome on different sample types. Machine learning algorithms is assessing antibody cross reactivity based on its binding signature and compared to its affinity.We have applied our technology to screen hundreds of antibodies from manufacturers. Please contact us to evaluate your antibodies. Selected antibodies are used subsequently for assay development.

Step4: Assay development

10+ years of expertise in multiplex assay development

Using diverse coupling techniques, we can immobilise many different types of molecules (antibodies, affimers, proteins, etc) and employ detection schemes (sandwich assay, single antibody, direct labeling etc) that fit your project specification. For standard bead-based ELISA schemes qualified antibodies are used for coupling, pairing, tuning and multiplexing. If you decide to send us samples (or sample are obtained from our biobank partners), our assay development service will include sample optimisation where we identify optimal dilutions for assay linearity and selection of assay buffers.

Step5: Assay Performance

Can the assay detect the analyte with sensitivity, precision and accuracy?

Following EMA, FDA, and CLSI’s guidelines we assess assay performance for Limits of Detection, Precision, Accuracy, and Linearity. You can then select our Biomarker Performance Service where your assays will be evaluated for their diagnostic performance (see optional Step6).

Step6: Biomarker Performance Service

Does your assay have diagnostic, prognostic, predictive power?

At this step we measure your samples and provide you with the raw data. Biomarker performance will then be assessed by our bioinformatics team using methods such as ROC, PCA, clustering, and advanced machine learning. Sub-panels of analytes or single hits with maximum diagnostic performance are identified and validated with clinical sample cohorts.

Step7: Assay Production & QC

ISO13485 compliant production service

Our production process that is compliant with the FDA & EMA guidelines guarantees that your assays will be produced with the best industry standards that can  meet IVD requirements. If needed, for large productions we can fully automate the process with our three robots. Manual production is also possible without any loss in quality. Our quality control extends the industry standards and includes proprietary QC methods that ensures the best assay precision on the market.