Collagen alpha-1(VI) chain (COL6a1) is a protein that in humans, the collagens are a superfamily of proteins that play a role in maintaining the integrity of various tissues. Collagens are extracellular matrix proteins and have a triple-helical domain as their common structural element. Besides, Integrin Alpha V (ITGaV) has been identified as an interactor of COL6A1, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human COL6a1 and recombinant human ITGaV. Briefly, COL6a1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul were then transferred to ITGaV -coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-COL6a1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant human COL6a1 and recombinant human ITGaV was shown in Figure 1, the EC50 for this effect is 0.19 ug/mL.