Fleet Bioprocessing Immunoassay Development and Bioconjugation
Fleet Bioprocessing is a UK-based contract research and development (R&D) and manufacturing provider specialising in the manipulation of antibodies and other proteins for pharmaceutical applications.
Working to meet the stringent quality standards of ISO 9001: 2015 and ISO 13485: 2016, Fleet’s areas of expertise include immunoassay development and bioconjugation applications.
The company uses its bioconjugation skills to label antibodies and other proteins with markers such as biotin, fluorescent dyes, enzymes and microparticles. These are often used as signal or capture reagents in immunoassays.
It also develops and executes immunoassays on platforms ranging from traditional enzyme-linked immunosorbent assay (ELISA) formats to the Gyrolab™ high-throughput system.
Development, qualification and execution of immunoassays
Fleet is experienced in the development of immunoassays on a range of platforms.
The company ensures that its customers’ assay requirements are clearly understood and documented in a product requirements document (PRD), which helps guide decision-making.
Fleet manages immunoassay development from the first stages of antibody selection. Guided by the company’s extensive supplier knowledge, the firm provides diligent screening of reliable commercial sources or generates new antibodies such as monoclonals, polyclonals and phage display antibodies.
Fleet combines antibodies in various assay architectures to yield a prototype immunoassay, including competitive or immunometric ELISA formats. Key variables such as antibody concentrations, conjugate incorporations and buffer formulations will be examined to establish optimal assay conditions.
Once the immunoassay design is finalised, Fleet conducts a series of qualification studies to confirm that the PRD requirements have been met.
European Centre of Excellence for the Gyrolab™ Immunoassay Platform
Fleet is appointed as the European Centre of Excellence for the Gyrolab immunoassay platform, which is widely used for pharmacokinetic (PK), pharmacodynamic (PD), host cell protein (HCP) and anti-drug antibody (ADA) assays.
The Gyrolab platform features rapid throughput and requires a very low sample volume, making it suitable for many pharmaceutical applications.
The system is based on a streptavidin-coated microfluidic solid phase in compact disc (CD) format with fluorescent detection. Fleet prepares biotin and fluorescent-labelled antibodies or antigens before optimising the Gyrolab assay based on the concentrations of these conjugates. The choice of assay affects aspects such as microfluidic sample volumes.
The Gyrolab platform is robust and predictable, typically allowing assays with high-analytical performance to be developed rapidly.
Clear establishment of the customer’s assay performance requirements, effective communication throughout the project, comprehensive documentation of the assay development and the final assay protocol are key factors in the success of Fleet’s approach to Gyrolab immunoassay development.
Bioconjugation and labelling of antibodies for immunoassay
Fleet’s dedicated team of chemists specialise in bioconjugation processes, including protein coupling methodologies and conjugate purification techniques.
Typical bioconjugation products include antibody or antigen labelled with biotin, fluorescent dyes, luminescent molecules, solid phases such as functionalised polystyrene / magnetic particles, and enzymes such as horseradish peroxidase (HRP) and alkaline phosphatase (AP), which can be harnessed into colorimetric, fluorometric or luminometric assay formats.
Optimal conjugate labelling can be a critical factor in obtaining high immunoassay performance. Fleet’s conjugate purification procedures ensure complete removal of high-molecular-weight antibody or antigen aggregate that can cause high background signals and hinder assay sensitivity.
Fleet also offers characterisation data confirming key conjugate characteristics such as colorimetric 4′-hydroxyazobenzene-2-carboxylic acid (HABA) / avidin test results to confirm biotin incorporation, or ultraviolet (UV) visible spectrophotometric data to confirm antibody concentrations and enzyme or dye incorporations.
Fleet’s characterisation data includes a Certificate of Analysis for each conjugate batch. To assure batch-to-batch reproducibility, Fleet understands and controls all key process variables within the conjugation process to ensure its conjugation methods are robust, reliable and reproducible in many applications.
Fleet regularly performs conjugations at scales from microgram to gram quantities of antibody or antigen.
About Fleet Bioprocessing
Fleet was founded in 2000 based on experience from diagnostic companies such as Amersham International, Kodak Clinical Diagnostics and Johnson & Johnson (Ortho-Clinical Diagnostics).
Its staff and consultants are experienced in the development of immunoassays for highly-regulated commercial applications and are knowledgeable in all aspects of assay design, validation, manufacture and troubleshooting.
Fleet provides a rapid response to customer enquiries and quick project execution. Complete the enquiry form on this page to speak to one of Fleet’s experts.
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Hampshire RG27 8DH