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PubMed1993

Validation of virus inactivation and removal for the manufacturing procedure of two immunoglobulins and a 5% serum protein solution treated with beta-propiolactone.

Intravenous immunoglobulins and serum protein solutions are manufactured from human plasma pools of healthy, screened donors. A step-by-step validation of virus removal and/or inactivation was performed for the manufacturing process, which includes cold ethanol fractionation, beta-propiolactone (beta-PL) treatment, UV irradiation, thermal inactivation and other chemical and physical purification steps. The total viral clearance factors achieved for the entire manufacturing process were by several magnitudes greater than the potential virus load of current plasma pools. Human immunodeficiency virus 1 (HIV-1) infectivity was reduced by > 13.4 log for 7S immunoglobulin, > 15.3 log for IGM enriched immunoglobulin and > 16 log for a 5% serum protein solution. In addition, high clearance rate for a broad spectrum of model viruses was demonstrated for all three blood derivatives being > 23.2 to > 27.8 log for pseudo rabies virus (PSR), > 12.3 to > 22.6 log for vesicular stomatitis virus (VSV) and 6.9-10.6 log for simian virus 40 (SV40). For the beta-propiolactone inactivation step Hepatitis C model viruses, e.g. equine arteritis virus (EAV) and bovine viral diarrhoea virus (BVDV) were also investigated.

Blood↗
PubMed1995

Evaluation by polymerase chain reaction on the effect of beta-propiolactone and binary ethyleneimine on DNA.

Inactivating treatments for viruses such as pasteurization or alkylation by beta-propiolactone or binary ethyleneimine were tested for their capacity to modify nucleic acids. The modification of a nucleic acid was measured as the decrease in spot intensity in Southern blots after polymerase chain reaction (PCR) amplification. The inactivating treatments were applied to cellular and viral genomic material from a human lymphoblastoid cell line immortalized by Epstein Barr Virus (EBV), which produced a monoclonal antibody. Pasteurization did not modify the ability to amplify and detect cellular or viral DNA. Binary ethyleneimine strongly reduced the amount of detectable DNA and beta-propiolactone under particular conditions of incubation abolished all trace of DNA.

Alkylating Agents↗
PubMed1981

Reactions of beta-propiolactone, beta-butyrolactone and gamma-butyrolactone with nucleic acids.

Reactivity of beta-propiolactone, beta-butyrolactone and gamma-butyrolactone with guanosine, RNA, DNA and 4-(p-nitrobenzyl)pyridine was studied. beta-Propiolactone was 50--100 times more reactive with all the nucleophiles than beta-butyrolactone whereas gamma-butyrolactone was completely inactive. The rate of alkylation by the lactones was guanosine greater than RNA = denatured DNA greater than double-stranded DNA. The type of the adducts formed were characterized by fluorescence and ultraviolet spectroscopy. Similar alkylation products were formed by the two lactones. The main sites alkylated were N-1 at adenosine, N-3 at cytidine and N-7 at guanosine. The results suggest that the carcinogenic potency of the lactones correlates with their reactivity rather than with specificity of the adducts formed.

4-Butyrolactone↗
PubMed1982

Clinical evaluation of the hepatitis safety of a beta-propiolactone/ultraviolet treated factor IX concentrate (PPSB).

The association of viral hepatitis, type B, with the use of prothrombin complex concentrates (PPSB) has been well documented. PPSB prepared from cold-sterilized plasma (beta-propiolactone treated and UV irradiated) has been shown not to induce hepatitis in chimpanzees. 500 U of cold-sterilized PPSB were infused into 5 healthy male volunteers. Previous to, and up to 6 months after, PPSB-application in addition to careful clinical investigations, the following hepatitis parameters were determined: SGOT, SGPT, y-GT, alkaline phosphatase, total serum bilirubin, HBsAg, HBcAb and HBsAb. On the basis of all of the above parameters there was no indication of induction of viral hepatitis following the application of PPSB prepared from beta-propiolactone/UV treated plasma.

Adult↗
PubMed1983

Evaluation of thrombogenicity of beta-propiolactone/ultraviolet (beta-PL/UV) treated PPSB in chimpanzees.

The thrombogenicity of beta-PL/UV-treated PPSB (factor IX concentrate) was evaluated in chimpanzees. PPSB isolated from beta-propiolactone-treated and UV-irradiated plasma was injected into chimpanzees at a dose of approximately 100 units/kg body weight. An FDA licensed PPSB preparation served as the negative control, and a preparation containing activated as well as precursor clotting factors served as the positive control. 15 minutes, 1 h, 4 h, and 24 h after the PPSB application the following parameters were determined in the chimpanzee blood: factors II, VII, IX, X, VIII, fibrinogen, AT III, thrombin coagulase, Quick value, APTT and platelet count. Neither the untreated control preparation, nor the PPSB isolated from beta-propiolactone-treated and UV-irradiated plasma, showed signs of thrombogenicity in the chimpanzee model. The positive control indicated that the chimpanzee is a suitable model for the thrombogenicity testing of activated clotting factors.

Animals↗
PubMed1991

Principles of selective inactivation of viral genome. V. Rational selection of conditions for inactivation of the viral suspension infectivity to a given extent by the action of beta-propiolactone.

The influence of the initial concentration of beta-propiolactone, the composition of the solution, temperature, and pH on the bacteriophage MS2 infectivity inactivation kinetics has been studied. Rate constants have been determined for the infectivity inactivation and for the change in the concentration (the consumption) of the reactant under inactivation conditions. These constants have been shown to permit a sufficiently precise description of the phage MS2 survival curves under the action of beta-propiolactone. These data have been used to put forward a kinetic approach for the rational determination of conditions for inactivation of the viral infectivity to a required extent with agents whose concentration decreases during inactivation as a result of hydrolysis and reactions involving the medium components.

Bacteriophages↗
PubMed1985

Induction of in vitro transformation by near-u.v. light and its interaction with beta-propiolactone.

The ability of u.v.-A light (320-400 nm) to induce cellular transformation in vitro and to modify chemical carcinogen-induced cellular transformation was investigated in BALB/c 3T3 cell cultures. When administered as a series of nontoxic exposures, u.v.-A alone was found to induce cellular transformation as a linear function of the numbers of u.v-A exposures. Possible interactions of u.v.-A with environmentally encountered chemical carcinogens were studied by examining the effects of u.v.-A light exposures on cellular transformation in cells exposed to the direct acting carcinogen, beta-propiolactone (BPL), an alkylating agent, with a standard initiation/promotion protocol. Twenty-four hours after a single treatment with 2.5 micrograms/ml of beta-propiolactone, cells were exposed to 3.0 kJ/m2 of u.v.-A light. U.v.-A exposures were repeated weekly for up to 5 weeks, after which cells were fixed, stained and dishes were scored for type III transformed foci. Weekly exposures to u.v.-A alone for 5 weeks induced approximately 3 foci/dish. Treatment with BPL alone induced approximately 1 focus/dish (background was 0.17 foci/dish). A combination of the two treatments resulted in a marked increase in the yield of transformed foci/dish, with the u.v.-A enhancement increasing with increasing numbers of exposures (approximately 10 foci/dish after a single exposure to BPL and five u.v.-A exposures). These results suggest a synergistic interaction between BPL and subsequent u.v.-A exposures in the induction of in vitro neoplastic transformation.

Animals↗
PubMed1962

Destruction of enteric bacteria in liquid egg with beta-propiolactone.

Liquid whole egg or egg white, inoculated with Escherichia coli 1485, Salmonella senftenberg ATCC 8400, or Salmonella typhimurium 84-I, was treated with concentrations of beta-propiolactone ranging from 0.05 to 0.3%. Egg white containing 1 x 10(3) to 1 x 10(6) cells of E. coli 1485 per ml was sterilized in 1 hr at 27 C by lactone concentrations of 0.2 and 0.3%. Egg white containing 1 x 10(5) cells of S. senftenberg ATCC 8400 per ml was sterilized in 12 hr at 10 C by 0.1% lactone and in 2 to 3 hr by 0.3% lactone at the same temperature. Liquid whole egg inoculated with 1 x 10(5) cells of either species of Salmonella was sterilized in 4 to 5 hr at 10 C with 0.2% lactone or in 2 to 3 hr by 0.3% lactone at this temperature. A mild heat treatment of either 15 min at 37 C or 1 min at 55 C markedly shortened the exposure times required for sterilization by beta-propiolactone at 10 C. After disinfection was complete, the lactone-treated liquid whole egg was reinoculated with low cell numbers of either species of Salmonella to determine the presence of residual lactone or toxic products. Liquid whole egg treated with 0.2% lactone would support the growth of salmonellae after 13 to 14 hr at 10 C. A heat treatment of 45 min at 37 C or 10 min at 55 C immediately after addition of 0.2% lactone allowed growth of the salmonellae in the lactone-treated liquid whole egg. No evidence of residual toxicity from the lactone treatment was found. The amount of lactone needed to prevent the outgrowth of low cell numbers of either strain of Salmonella in liquid whole egg was quantitated. Liquid whole egg containing 0.06 to 0.07% lactone would not support salmonellae growth from inocula of 1 to 10 cells per ml of egg. Lactone concentrations above 0.08% prevented outgrowth of salmonellae inocula of 10 to 200 cells per ml of liquid whole egg.

Anti-Infective Agents, Local↗
PubMed1976

Gas chromatographic trace analysis of beta-propiolactone in sterilized serum proteins.

A method for detecting traces of beta-propiolactone, which was previously described by Schmitz-Masse, has been modified and adapted to the analysis of protein solutions. Using this method, the rate of hydrolysis of beta-propiolactone (beta-P1) was examined and beta-P1 sterilized commercial products were analyzed for possible residual nonhydrolyzed beta-P1.

Blood Proteins↗
PubMed1975

[Beta-propiolactone for cold sterilization of heart valve transplants].

Biologic cardiac valve substitutes were sterilized at room temperature using beta-propiolactone with a 0.5 vol.% solution within 120 minutes, with a 1.0 vol.% solution within 90 minutes, and with a 1.5 resp. 2.0 vol.% solution within 60 minutes. The physico-chemic properties of the graft can be influenced by appropriately chosen concentration and sterilizing period. Enlarged procedures for decontamination are not necessary because of the rapid dissolution of aqueous beta-propiolactone solutions.

Animals↗
PubMed1976

[Collagen synthesis, studied by sub microscopic methods in beta-propiolactone and in tendon homografts, preserved by gamma irradiation].

In dog experiments homologous tendos conserved in beta-propiolacton and by means of gamma-radiation have been transplanted by the authors. After the 1st, 3rd and 8th post-operative week various regions of the graft and the recipient tendon stump have been examined by means of light--, polarisation microscopic and electron microscopic method. The examinations have unanimously shown that the tendon grafts conserved in beta-propiolacton are reorganized quickly, within 8 weeks nearly completely besides hardly visible inflammatory reaction. In the meantime, the originally collagen fibrous framework is decomposed and on it a new collagen fibrous structure develops, and thus a new tendon--capable to function--is formed. On the other hand, the reorganization of the grafts conserved with gamma-radiation is protracted or it fails to occur, and cicatrization is observed. Some problems of the modern tissue preservation, the collagen synthesis occurring in the active fibroblasts, the secretion of the collagen precursory substance into the extracellulary space, as well as the formation, the structure and the constitution of the newly synthesized collagen fibres--forming the transplant--are discussed by the authors. The obtained results are demonstrated in light-, polarization microscopic and electron microscopic pictures.

Animals↗
PubMed1980

[Tolerance of beta-propiolactone-treated and UV-irradiated rat serum].

beta-Propiolactone-treated and UV-irradiated rat serum was injected into rats to investigate its ability to generate new immunogenic determinants. No inflammatory/cytotoxic reaction was observed and antibodies directed against the beta-propiolactone-treated and UV-irradiated serum could not be detected.

Animals↗
PubMed1986

Pyrene acts as a cocarcinogen with the carcinogens benzo[a]pyrene, beta-propiolactone and radiation in the induction of malignant transformation in cultured mouse fibroblasts; soybean extract containing the Bowman-Birk inhibitor acts as an anticarcinogen.

Pyrene was found to act as a cocarcinogen in the induction of transformation of cultured Balb/c3T3 cells by three different types of carcinogens: a direct acting chemical carcinogen, beta-propiolactone, a chemical carcinogen requiring metabolic activation, benzo[a]pyrene, and a physical carcinogen (60Co) gamma radiation. Since pyrene enhanced transformation in vitro by approximately the same amount for all the carcinogens tested, these results suggest that the carcinogenic action of pyrene is not related to carcinogen metabolism or uptake in vitro. An extract of soybeans containing the Bowman-Birk protease inhibitor was shown to reduce transformation induced by beta-propiolactone, benzo[a]pyrene and gamma-rays, both with and without the cocarcinogenic effect of pyrene, to background levels; the magnitude of the reduction in transformation by the protease inhibitor preparation was unrelated to the concentration of carcinogen. Neither the mechanism for the cocarcinogenic action of pyrene not the anticarcinogenic effect of the soybean extract is known, but several hypotheses are discussed.

Animals↗
PubMed1971

Effect of -propiolactone on Sendai virus.

The biological properties (infectivity, hemagglutination, hemolysis, cell fusion, neuraminidase) of Sendai virus were dissociated on the basis of sensitivity to beta-propiolactone, by freeze-thawing, by heating at different temperatures, and by adsorption-elution with formalinized chicken erythrocytes. Possible mechanisms whereby beta-propiolactone selectively destroys viral infectivity are discussed.

Alkylating Agents↗
PubMed1972

Immunogenicity of rabies virus inactivated by -propiolactone, acetylethyleneimine, and ionizing irradiation.

Ionizing radiation, beta-propiolactone, and acetylethyleneimine were compared for their ability as virus-inactivating agents for the preparation of rabies vaccine. Each agent reduced viral infectivity exponentially; ionizing radiation also destroyed viral hemagglutinin. The vaccine prepared by ionizing radiation was equal or superior to that prepared by beta-propiolactone in its ability to protect mice from rabies infection. The acetylethyleneimine-treated vaccine was a less potent immunogen.

Acetates↗
PubMedDate not supplied

[Fibrin removal from mouse immune ascitic fluids to arbovirus using beta-propiolactone].

A modification of acid precipitation of fibrin in immune ascitic fluids (IAF) employing beta-propiolactone has been developed. The procedure removes fibrin completely without affecting the titres of specific antibody. This modification is not inferior to previously described methods for fibrin removal from IAF and has a number of advantages. In addition to removing fibrin from IAF, beta-propiolactone prevents possible virus contamination of IAF and sterilizes the preparation additionally.

Antibodies, Viral↗
PubMed1985

Inactivation of the Hutchinson strain of non-A, non-B hepatitis virus by combined use of beta-propiolactone and ultraviolet irradiation.

A beta-propiolactone/ultraviolet irradiation procedure (beta PL/UV) has been evaluated for its ability to inactivate 30,000 chimpanzee infectious doses of the Hutchinson strain of non-A, non-B (NANB) virus. The chimpanzees were inoculated with plasma to which this dose of the titrated virus had been added prior to application of the beta PL/UV process in accordance with a procedure used for licensed blood derivatives in Germany. Neither animal developed hepatitis. When subsequently challenged with the same contaminated plasma, which had not been sterilized, both animals promptly developed typical NANB hepatitis. This study extends the high (approximately 10(7)-fold) process efficiency of the beta PL/UV procedure previously reported for hepatitis B virus to a blood-borne NANB virus.

Animals↗
PubMed1988

Inactivation of the Hutchinson strain of hepatitis non-A, non-B virus in intravenous immunoglobulin by beta-propiolactone.

beta-propiolactone (beta-PL) treatment has been evaluated for its ability to inactivate 10(3.5) chimpanzee infectious doses (CID50) of the Hutchinson strain of hepatitis non-A, non-B virus (HNANBV). Two chimpanzees were inoculated with a beta-PL-treated immunoglobulin solution to which this dose of the titrated virus had been added prior to beta-PL treatment. beta-PL treatment was performed in accordance with the production procedure used for a licensed intravenous immunoglobulin preparation. Neither animal developed hepatitis. When subsequently challenged with the same spiked immunoglobulin solution that had not been beta-PL treated, both animals developed clear-cut hepatitis non-A, non-B. The results of this experiment demonstrate that beta-PL treatment is effective for the inactivation of hepatitis non-A, non-B virus in intravenous immunoglobulin.

Animals↗