The present invention provides enrichment, detection methods and applications of GPC-3 complexes in blood samples. The enrichment method includes A. collecting
peripheral blood from
liver cancer patients, placing it in an
anticoagulant tube and mixing the
whole blood; B. adding multiple volumes of
red blood cell lysis solution, pipetting and mixing, lysing on ice, centrifuging and discarding the red supernatant. If the
cell pellet is still red, repeat this step; C. washing the pellet 1-2 times, adding PBS or normal
saline to resuspend the pellet, centrifuging and discarding the supernatant; D. counting white blood cells, adding PBS to resuspend the
cell pellet, taking an appropriate amount to fill the
pool, letting it stand for 2-3 minutes, and manually counting the white blood cells using the four large squares in the four corners of the low-power
microscope. The calculation formula is as follows: WBC / L=N / 4*10 5 *10 4 , and calculate the number of white blood cells per milliliter of the original suspension, WBC / ml original suspension = N / 4*10 4 *
Dilution factor; E. Add
lysis buffer containing 1%
protease inhibitor and 1%
phosphatase inhibitor to the obtained cells, refrigerate and let stand for a certain period of time, and then
centrifuge to obtain the supernatant as the sample to be tested to achieve GPC-3 enrichment.