Background: In order to detect anti-HPV16E7 antibody in serum, a highly sensitive and rapid detection method based on chemiluminescence immunoassay and immunomagnetic separation was introduced. The technique that was developed is a novel, sensitive chemiluminescence (CL) immunoassay for HPV16E7 antibody detection.
Methods: Balb/C mice were immunized with HPV16E7 fusion protein to prepare monoclonal antibody against HPV16E7. The biotinylated antigen was prepared as immunomagnetic beads and its stability was tested (IMBs). The protocol used horseradish peroxidase (HRP) labeled HPV16E7 antigen and immunomagnetic beads (IMBs). The antibody induced the formation of IMBs-mAbs HPV16E7-HRP labeled antibody structures. IMBs were applied to capture CEA and immobilize CEA through the external magnetic field. Oxidized luminescence substrate can be catalyzed by HRP on antibody surface to generate optical signals which were detected by luminometer.
Results: HPV16E7 monoclonal antibody was prepared and validated. The HPV16E7 antigen can efficiently bind to the bead with a conjugation rate of 72%. The biological activity of IMBs did not decrease significantly when stored in the dark at 4℃ for 2 months. The sensitivity and stability of this proposed method were excellent and showed a good linear relationship (Y = 1.3203 X + 0.7, R2 = 0.9952).
Conclusions: This proposed technique showed excellent performance in quantitative measurement of HPV6E7 and was expected to be used in clinical detection.