Test principle of the Dip’n’Check

How do the AAV Dip'n'Check Tests work?

Lateral flow tests are based on the passing of a liquid sample through a series of capillary beds driven by capillary forces. The analysis of the presence of a specific substance in the sample follows the same principal as the affinity chromatography. 

The surface of the tests contains reactive molecules that show either a positive or negative visual result depending on the presence or absence of the substance. Lateral flow tests generally use specific conjugates usually consisting of an antibody specifically binding the target molecule labelled with a visual tag, e.g. colloidal gold.

The samples to be analyzed are pre-incubated with the conjugates before the application of the sample to the test strip. 

 After the conjugate binds to the target molecule, the complex migrates further along the test strip until reaching the test line containing an immobilized reagent to capture the target-conjugate complex. With the binding of the complex, the visual tag accumulates at the test line and shows a visual change. 

In the case of the Dip´n´Check AAV a purple-red color confirms the presence of the target molecule. In contrast to the test line, the control line should appear independently of the presence of the target molecule to confirm the validity of the test.

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The AAV sample is pre-incubated with two differently conjugated AAV antibodies to allow coupling of the gold conjugate and for immobilization at the test line. The two antibody solutions contain the same antibody but labelled with two different conjugates. 

The antibodies used for the Dip'n'Check AAV kits are AAV particle antibodies exclusively binding to fully assembled AAV capsids. Both AAV antibody conjugates delivered with this test are mixed in equal volumes with running buffer and the AAV sample, and incubated for 10 min to allow binding of the antibodies. 

The test strip is then placed in the solution to allow complex formation with the gold nanoparticles at the conjugate pad and further migration by capillary force to the test and control lines. 

The Dip´n´Check AAV quick test is a semi-quantitative assay with a detection range of 1.0E+08 – 1.0E+10 capsids. Within this range, a 2-fold concentration difference can clearly be detected. There is no difference in the antibody affinity to empty and full capsids.

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