Development of a device for the fast analysis of a high number of biofluids samples containing hormones bound to proteins (GPG.6038)

Biochemical analytical methods to study variations in contents of hormones, drugs and proteins in body and other biological fluids to be applied in off-laboratory settings. We explored and developed:

• A prototype of a lateral flow immune analysis flow (LFIA) method for progesterone in cow's milk. Important issues were how to pre-purify the milk with relatively simple methods and to achieve the desired sensitivity.
• The ARIS method for progesterone. This method seems not suitable for re-activation with the FAD-progesterone conjugate and appears too vulnerable to the composition of the biomatrix and did therefore not work with fresh cow milk.
• A minimally invasive sampling and storage technique, allowing to sample time-profiles of a wide variety of analytes over periods up to 4 days in freely moving subjects, including human, cow and rats.
• Interfaces to connect the sampling device to detection devices including micro-HPLC and MS for the analysis of time-profiles.
• A miniaturized flow-reaction device allowing to measure in-line enzymes and analytes such as glucose oxydase, lactate oxydase, glucose and lactate, down to sub-femto-mole amounts.

Utilization
A prototype LFIA method, that can now be optimized for clinical and veterinarian use.
The micro-reactor is a robust device for studying enzyme and receptor kinetics under both physiological and severe non-physiological conditions with a minimum amount (down to sub-micrograms or femto-molar range).
The microfiltration devices allow sterile sampling of blood and other biological fluids over periods as long as 96 hours, both bed-side and from freely active subjects (e.g. to be applied in the general practice). Microfiltration allows also continuous sterile sampling from bioreactor vessels.
The continuously collected and stored microfiltration samples remain stable for several days and can be sent to any location by mail for analysis. The proposed technologies may assist biochemical analysis outside the biomedical field.