Subject(s)
Hemostasis , Peptides , Amino Acid Sequence , Anticoagulants/therapeutic use , Antithrombin III/analysis , Biological Assay , Factor IX/analysis , Factor V/analysis , Factor VII/analysis , Factor VIII/analysis , Factor X/analysis , Factor XI/analysis , Factor XII/analysis , Factor Xa , Female , Fibrinolysin/analysis , Glycoproteins/analysis , Heparin/therapeutic use , Humans , Kinetics , Male , Monitoring, Physiologic , Partial Thromboplastin Time , Plasminogen/analysis , Platelet Factor 3/analysis , Platelet Factor 4/analysis , Prekallikrein/analysis , Protein C , Prothrombin/analysis , Prothrombin Time , Thrombin/analysisABSTRACT
Hepatitis B virus infectivity can be removed from human plasma used to prepare a gamma-globulin product by ion-exchange chromatography with commercially available resins. A pooled human plasma sample free of hepatitis markers was contaminated with 10,000 chimpanzee infectious doses of hepatitis B virus per milliliter from a known infectious inoculum. The contaminated plasma was diluted to yield a weakly positive radioimmunoassay result for HBsAg and then processed over a double-column chromatography system containing anionic ion-exchange resins to yield highly purified gamma-globulin. Two chimpanzees were each inoculated intravenously with this product while a control animal received non-treated material. The 2 experimental animals each received a potential challenge of 3,000 ID and showed no serologic evidence of the disease during 9 months of evaluation. The control animal received 100 ID and developed hepatitis B 25 weeks after the challenge.
Subject(s)
Drug Contamination/prevention & control , Hepatitis B virus/isolation & purification , gamma-Globulins/adverse effects , Animals , Chromatography, Ion Exchange , Female , Hepatitis B/etiology , Hepatitis B/prevention & control , Hepatitis B Antibodies/biosynthesis , Hepatitis B Surface Antigens/analysis , Humans , Male , Pan troglodytes , Safety , gamma-Globulins/isolation & purificationABSTRACT
Utilizing a unique human plasma sample that contained extremely high concentrations of hepatitis B surface antigen (HBsAg), we were able in the laboratory to rapidly screen the binding efficiency for HBsAg of various ion-exchange resins that can be used to purify gamma globulin product from human plasma. 'Nonrigid' resins like QAE-Sephadex and DEAE-Sephadex showed a high affinity for HBsAg by removing greater than 10(4) of applied challenge. Much lower binding was achieved by 'rigid' resins like DEAE-Sepharose CL-6B and DEAE-Spherodex. Also examined were the usefulness of certain auxiliary procedures, special filters and resins, that offer the potential to remove HBsAg. We failed to find any significant removal of the HBsAg, and in some cases large losses of the gamma globulin product occurred. While encouraged by our findings, certain precautions are discussed concerning the conversion from large-scale production of gamma globulin by the conventional alcohol method to one utilizing chromatographic techniques.
