Study On The Methods Of Human Immunoglobulin-Specific Purification And Separation

Separation and purification of human hepatitis B immunoglobulin (HHBIG) and human rabies immunoglobulin (HRIG) were studied in this study, and in order to resolve the problem that low-ligand density. about affinity medium which were made of biological macromolecules such as the third affinity igand, we discussed seriously. Main results are as follows:(1) Through the analysis of HHBIG materials, affinity chromatography (AC) were confirmed the optimal chromatography. We made an affinity medium which was used to purifying HHBIG, and the affinity medium was named Sepharose 4FF-HHBIG, and the operating conditions of AC were optimized. For the materials which titer was 95 IU/mL, the purification fold was 30 and the recovery of HHBIG activity was 45%; for the materials which titer was 45 IU/mL, the purification fold was 40 and the recovery of HHBIG activity was 50%. Purification chaperon PEG D (2%, w/w) was screened out and applied in the purification of HHBIG, the recovery of HHBIG activity was increased 10-15% more than none PEG.(2) We made an affinity medium which was used to purifying HRIG. The affinity medium was named Sepharose 4FF-HRIG, and the operating conditions of AC were optimized. The results showed that the purification fold was 8 and the recovery of HRIG activity was 18%.(3) Modified PS microsphere was used as the affinity support and the third affinity igand was emplyed as the ligand, we made an affinity medium which was used to purifying HRIG. The affinity medium was named mPS-HRIG. Sepharose 4FF-HRIG affinity medium and mPS-HRIG affinity medium on purification efficiency and their physicochemical properties were compared. The results revealed that the ligand coupled to modified PS microsphere is about 2 times of that coupled to Sepharose 4FF; furthermore, the purification fold and recovery of HRIG activity of mPS-HRIG affinity medium higher than Sepharose 4FF-HRIG affinity medium, and mPS-HRIG affinity medium has higher mechanical strength. Therefore, mPS-HRIG affinity medium is an ideal affinity medium for HRIG purification.