Biochip assembly and assay method thereof

Abstract

The present invention is directed to a biochip assembly comprising a semi-permeable membrane and an assay method using said biochip assembly for carrying out cell based assays.Ideally, such a method involves measuring the migration of cells in a channel under the influence of an analyte wherein said cells are separated from said analyte by a semi-permeable membrane and said analyte and / or said cells are subjected to controlled flow conditions.

Description

FIELD OF THE INVENTION[0001]The present invention is directed to a biochip assembly comprising a semi-permeable membrane and a method using said biochip assembly for carrying out cell based assays.BACKGROUND OF THE INVENTION[0002]The ability to monitor migration of biological cells through tight layer of other cells and tissues is crucial for understanding of mechanism of many life threatening diseases and development of modern therapeutic drugs. This migration is typically triggered by the presence of a particular chemical either immobilized on a surface or diffused through a tissue.[0003]In inflammatory conditions, for example, the migration of leukocytes from blood vessels into diseased tissues is crucial to the initiation of normal disease-fighting inflammatory responses. At the same time, this process, known as leukocyte recruitment, is also involved in the onset and progression of debilitating and life-threatening inflammatory and autoimmunne diseases. Thus, the ability to con...

AI Technical Summary

Benefits of technology

[0047]Ideally, the migration of cells is transmigration and the method of the invention facilitates the transmigration of cells through the semi-permeable membrane.

Problems solved by technology

However, margination is not commonly accepted as yet in the three step migration process described above.

Although considerable effort has been invested on the study of lymphocyte chemoattractants, they remain poorly characterized relative to monocyte and neutrophil chemoattractants.

However, no molecule identified primarily as a lymphocyte chemoattractive factor has been sequenced and cloned.

This excessive time span is necessary in order to get reasonable statistics of cell migration.No physiological flow can be established in this assay thus is not possible to monitor cells though all stages of leukocyte recruitment.There is no control of the gradient: chemokine diffusion in the body might be different than in vitro as it takes longer to get a cell migration on in vitro assays.Changes in cell morphology during chemotaxis cannot be observed in real-time (because cells transmigrate through the filter).

In addition, Boyden chamber assays cannot readily answer many questions related to the leukocyte migration.

However, in blood even a short persistence of a soluble chemokine gradient is not feasible because the constant flow of plasma removes the soluble chemokines from the site of their production.

However, many of these cells are difficult to grow in vitro and, when grown, are not morphologically similar to in vivo tissue.

However, animal studies are costly, time-consuming and are limited, by throughput, to characterize no more than a few compounds.

Furthermore, several drugs have shown unanticipated or unpredicted side effects only after reaching clinical trials or wide-scale release to the public.

The complexity and interplay of biological processes that must be simulated to predict the ADMET properties of a compound far exceed the capabilities of currently available methods and tools.

Even if these biological functions could be faithfully reproduced in vitro, a difficulty remains in getting the capacity and format of the assay to facilitate testing and analysis of thousands of compounds.

However, there are several disadvantages when using the parallel chamber.

For example, the parallel flow chamber requires a substantial amount of the drug candidate for the experiment.

Furthermore, setting up the experiment is often time consuming and rather complex.

Efficient metabolism of a candidate drug by a CYP450 enzyme may lead to poor pharmacokinetic properties, while drug candidates that act as potent inhibitors of a CYP450 enzyme can cause undesirable drug-drug interactions when administered with another drug that interacts with the same CYP450.

Consequently, such earlier-available, reliable pharmacokinetic information may result in greatly reduced drug development costs and / or increased profits from earlier market entrance.

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