Methods for inactivating pathogens using broad-spectrum pulsed light

A pathogen and pulse technology, applied in the field of inactivating pathogens with broad-spectrum pulsed light, can solve the problems such as BPL not raised

Inactive Publication Date: 2002-04-10
PUREPULSE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While BPL has been shown to be useful for the inactivation of various microorganisms on solid surfaces and / or within certain relatively clear liquids, it has not been suggested that BPL can be used to remove bioderivatives containing proteins and / or other biomolecules of interest. contamination in the composition

Method used

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  • Methods for inactivating pathogens using broad-spectrum pulsed light
  • Methods for inactivating pathogens using broad-spectrum pulsed light
  • Methods for inactivating pathogens using broad-spectrum pulsed light

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Inactivation of SV40

[0059] African green monkey kidney (AGMK) cells were inoculated with simian virus 40 (SV40, ATCC VR-305). Virus was harvested when 75%-100% of cells exhibited cytopathic effect (CPE). The virus stocks were frozen and stored at -60°C in culture medium containing 10%-15% fetal bovine serum (FBS). SV40-containing samples were titrated with AGMK cells. The culture medium used to grow and maintain AGMK cells is Eagles minimal essential medium (E-MEM), containing 5%-10% heat-inactivated FBS, 10 mg / ml gentamicin, 100 units / ml penicillin, and 2.5 mg / ml amphotericin B.

[0060] Cell cultures were grown at 36-38°C in a humidified 5-7% CO2 atmosphere.

[0061] SV40 in E-MEM containing 10% heat-inactivated FBS was added in a volume of 0.2 ml to sterile polyethylene sample containers. Twelve samples were prepared. Nine samples were treated with BPL. Three samples were treated with one pulse of light, three samples were treated with two pulses...

Embodiment 2

[0065] Example 2: Inactivation of HIV-1

[0066] CCRF-CEM cells were inoculated with HIV-type I strain HTLVIIIB (Vanderbilt University, Nashville, TN). HIV-1 was collected when the infectivity reached a minimum of 80% as determined by immunofluorescence assay (IFA). Frozen virus stocks, stored at -60°C in culture medium containing 10%-15% FBS.

[0067] HIV-1 containing samples were titrated with MT-2 cells (NIH AIDS Research and Reference Reagent Program. Cat #237). The medium used for the growth and maintenance of MT-2 cells was supplemented with 2 mM L-glutamine, 25 mM Hepes, 2 g / L NaHCO 3 , RPMI 1640 (with phenol red) in 15% heat-inactivated FBS and 50 mg / ml gentamycin. at 36-38°C in humidified 5-7% CO 2 Cell cultures were grown in the atmosphere.

[0068] 0.2 ml of HIV formulated in RPMI containing 15% heat-inactivated FBS was added to a sterile polyethylene sample container. Twelve samples were prepared. Nine samples were treated with BPL. Among them, three copies...

Embodiment 3

[0073] Example 3: Inactivation of CPV

[0074] A-72 cells (ATCC CRL 1542) were inoculated with CPV (ATCCVR-2017). Virus was harvested when 75%-100% of the cells showed cytopathic effect (CPE). Frozen virus stocks, stored at -60°C in culture medium containing 10%-15% FBS.

[0075]CPV-containing samples were titrated with A-72 cells. The medium used for the growth and maintenance of A-72 cells was E supplemented with 5%-10% heat-inactivated FBS, 10 mg / ml gentamicin, 100 units / ml penicillin, and 2.5 mg / ml amphotericin B. -MEM. Humidified 5-7% CO at 36-38°C 2 Cell cultures were grown in the atmosphere.

[0076] 0.2 ml of CPV in E-MEM containing 5% heat-inactivated FBS was added to a sterile polyethylene sample container. Twelve samples were prepared. Nine samples were treated with BPL. Among them, three copies were treated with one light pulse, three copies were treated with two pulses, and three copies were treated with three pulses. Three untreated samples served as con...

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Abstract

Methods for reducing the pathogen content of a biologically derived composition by illuminating the composition with at least one high-intensity, short duration pulse of incoherent polychromatic light in a broad spectrum. In the resultant treated compositions, biomolecules of interest remain biologically active. The content of pathogens, such as viruses, bacteria, pyrogens, fungi and / or prions, present in a biologically derived composition, such as blood serum, blood plasma or other blood product, including, insulin, transferrin, heparin, collagen, Factor VIII and / or Factor IX, or a composition containing monoclonal antibodies, proteins from genetically engineered cell lines or the like, is reduced by illumination with broad-spectrum pulsed light. The content of HIV-1, SV40, canine parvovirus or bovine viral diarrhea virus in a biologically derived composition is dramatically reduced with a single pulse of broad-spectrum light.

Description

Background of the invention [0001] The use of biologically derived compositions is ubiquitous in scientific research and in the manufacture of pharmaceuticals / therapeutics. Human blood is routinely used as a source of protein preparations for the treatment of various diseases and abnormalities; for example, fractions of plasma, providing factor VIII, factor IX, antithrombin, transferrin, albumin, immunoglobulins, and platelets , and other therapeutic agents. Tissue culture methods are commonly used in the production of many pharmaceutical / therapeutic agents and related compositions, such as recombinant DNA and / or recombinant proteins from genetically engineered cell lines, viral vectors, amino acids, peptones, insulin and monoclonal antibodies. Similarly, animal-derived reagents such as bovine serum albumin (BSA) and sheep blood fractions are routinely used in research and manufacturing. [0002] Since these compositions are derived from living organisms, including humans, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/04A61K35/12A61K35/14A61K35/76A61K38/00A61K38/16A61K38/17A61K38/28A61K38/43A61K39/395A61L2/00A61L2/08
CPCA61L2/0011A61L2/0029A61L2202/11
Inventor H·W·科弗J·M·伯格尔C·J·麦克唐纳德S·马洛伊A·H·布什内尔J·R·库珀
Owner PUREPULSE TECH
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