جداسازي و خالص سازي كمپلكس آلفا آنتي كيموتريپسين آنتي ژن ويژه پروستات با استفاده از روش هاي كروماتوگرافي

Separation and purification of prostate specific antigen - a-antichymotrypsin complex from blood serum using various chromatographic procedures

In this research, we explored the various chromatographic procedures with the intention of preparing pure prostate specific antigen - a-antichymotrypsin complex (PSA-ACT) from human blood serum. We found that in pH=7.2, both free PSA and PSA-ACT molecules with negative charge are bound to the column containing DEAE-Sephacel. They could be separated from other proteins by applying a change to linear gradient of NaCl at pH=7.2. A large quantity of Albumin in serum was separated from free PSA and PSA-ACT molecules by Con A-Sepharose column, but the PSA compounds in free and complex form were not separable in this stage. In the last stage, by taking the advantages of large difference in molecular size between free PSA and PS A-ACT complex and applying gel filtration chromatography and using a column containing Sephacryl S-200, the pure PSA-ACT complex attained without other contaminating serum proteins. Finally it is appeared that the combined use of these chromatographic procedures would permit the isolation of pure PSA-ACT complex from human serum that could be used in the preparation of more specific antibodies and suitable calibration of a new generation of immunoassay for PSA analysis.