Purification of Alpha-Fetoprotein from Human Umbilical Cord Plasma

Alpha-fetoprotein (AFP) is a sera protein that involves in fetal development, produced by the fetal liver and yolk sac. Human AFP (hAFP) is not detected in adults but it is increased in cases of hepatocellular carcinoma (HCC), cirrhosis and other cancers like germ cell cancer, stomach and intestine cancer. hAFP has been used as an important marker for HCC serological diagnosis and it has recently become a target for HCC immunization studies. Umbilical cord blood has approximately 150–250 μg/ml hAFP but it is usually discarded in hospitals. In order to take advantage of human umbilical cord blood as hAFP nature material, this study have established a three-step chromatography process for hAFP purification including removal of albumin by Hitrap blue HP column, hAFP captured chromatography using rabbit anti-hAFP IgG column, and filtration chromatography through Sephacryl S100 HR column. By native-PAGE, western blotting and bradford assays, the hAFP purity was showed more than 95% and 80 μg of hAFP per mL of initial cord plasma was recovered. Our established hAFP purification process can be performed on a larger scale for later applications in the serodiagnosis of HCC and other hAFP related diseases.