Purification of Hyaluronan Binding Proteins from Human Normal and Cancer Serum

Background: Analysis of human cancer serum has revealed the presence of high amounts of hyaluronan (hyaluronic acid, HA) when compared to human normal serum, It is well documented that HA and its receptors, known as hyaladherins (HABPs) are involved in matrix regulation, cell proliferation, migration and at the matrix proper but also with hyaluronan at the plasma membrane as a cell surface receptors and thus influence cell physiology including secretion of this protein into the circulatory system. Methods: Normal serum and colon cancer serum samples were included in this study using biochemical techniques such as gel permeation, strong anion exchange chromatography, single dimension electrophoresis and western blot analysis. Results: This study is based on the clinical work of normal serum and colon cancer serum. The description of the procedure was given for the fractionation of serum proteins mainly HABPs from 20 normal and 15 colon cancer patients by using special biotinylated hyaluronan probe. Conclusion: To evaluate whether serum HABPs levels could be used as diagnostic marker for human cancer. The semi purified serum from normal and colon cancer patients showed mainly a major protein (57kDa) and a minor one (30kDa) by overlay experiments with b-HA probe and these results were confirmed by competition experiments with cold HA.