US20150210986A1 - Virus purification and formulation process - Google Patents

Virus purification and formulation process Download PDF

Info

Publication number
US20150210986A1
US20150210986A1 US14/423,394 US201314423394A US2015210986A1 US 20150210986 A1 US20150210986 A1 US 20150210986A1 US 201314423394 A US201314423394 A US 201314423394A US 2015210986 A1 US2015210986 A1 US 2015210986A1
Authority
US
United States
Prior art keywords
virus
purification
alum
disclosed
filtration
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US14/423,394
Other languages
English (en)
Inventor
John J. Vicalvi, JR.
Edward G. Hayman
Joseph Makowiecki
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Global Life Sciences Solutions USA LLC
Original Assignee
GE Healthcare Bio Sciences Corp
Xcellerex Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by GE Healthcare Bio Sciences Corp, Xcellerex Inc filed Critical GE Healthcare Bio Sciences Corp
Priority to US14/423,394 priority Critical patent/US20150210986A1/en
Publication of US20150210986A1 publication Critical patent/US20150210986A1/en
Assigned to XCELLEREX INC. reassignment XCELLEREX INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: HAYMAN, EDWARD, MAKOWIECKI, JOSEPH, VICALVI, JOHN
Assigned to GE HEALTHCARE BIO-SCIENCES CORP. reassignment GE HEALTHCARE BIO-SCIENCES CORP. MERGER (SEE DOCUMENT FOR DETAILS). Assignors: XCELLEREX, INC.
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/24011—Flaviviridae
    • C12N2770/24111—Flavivirus, e.g. yellow fever virus, dengue, JEV
    • C12N2770/24151—Methods of production or purification of viral material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/24011—Flaviviridae
    • C12N2770/24111—Flavivirus, e.g. yellow fever virus, dengue, JEV
    • C12N2770/24161—Methods of inactivation or attenuation
    • C12N2770/24163—Methods of inactivation or attenuation by chemical treatment

Definitions

  • the present invention relates generally to a method of purifying biologics, such as a virus or modified virus, and a method of formulating the biologic, for example, a virus and adjuvant.
  • biologics such as a virus or modified virus
  • a virus purification process for producing highly purified virus product having no residual sucrose without significant virus loss such as that which is typically encountered by prior art virus purification processes.
  • the disclosed process captures and purifies the virus, separating it from the host cell proteins and DNA, and leaving the host cell proteins and DNA behind.
  • the process also can be used to inactivate and/or concentrate the virus sufficiently for use in formulations.
  • Also disclosed herein is a one-step process for producing a formulation wherein the concentrated virus particles are not aggregated or clumped together.
  • the disclosed process utilizes adjuvants and buffer exchanges to process the virus, and results in a final, buffered virus formulation.
  • An embodiment of the process can also be used for formulation of other biologics.
  • FIG. 1 is a process chromatogram for Experiment Y1626A, an embodiment of the new downstream process disclosed herein that is used to purify and formulate a harvest of modified YF virus.
  • FIG. 2 is a process chromatogram for Experiment Y1632A, an embodiment of the new downstream process disclosed herein that is used to purify and formulate a harvest of modified YF virus, comparing elutions using CELLUFINE® sulphate and CAPTOTM Core 700.
  • FIG. 3 is a process chromatogram for Experiment Y1637A.
  • FIG. 4 is a process chromatogram for Experiment Y1639A using CELLUFINE® Sulfate column.
  • FIG. 5 is a process chromatogram for Experiment Y1639A using CAPTOTM DeVirS Column.
  • FIG. 6 is a flow diagram for an embodiment of the old downstream purification process and a flow diagram for an embodiment of the new downstream purification process.
  • FIG. 7 is a flow diagram for the embodiment of the old downstream purification process shown in FIG. 6 and a flow diagram for an embodiment of the new and improved downstream purification process.
  • FIG. 8 is a flow diagram for another embodiment of an old downstream purification process and a flow diagram for an embodiment of the new and improved downstream purification process, Number 3.
  • the inventors of the present subject matter have now discovered a process for preparing a highly purified biological composition, such as, for example, a virus.
  • the process is particularly useful for purifying a flavivirus, e.g., a Yellow Fever Virus, Japanese Encephalitis virus, Dengue virus, and West Nile virus.
  • a flavivirus e.g., a Yellow Fever Virus, Japanese Encephalitis virus, Dengue virus, and West Nile virus.
  • the examples provided herein are for purifying a Yellow Fever virus, but with no more than routine experimentation, could be used to purify other types of viruses.
  • the disclosed process significantly reduces the loss of viral particles as compared to prior art methods for obtaining a highly purified sample of a biologic.
  • Use of an embodiment of the disclosed method also significantly lowers the amount of sucrose in the final purified product as compared to prior art methods of purification.
  • YF Yellow Fever
  • YF virus may be grown on Vero cells, harvested, inactivated, and purified.
  • prior art methods for downstream purification of a virus provide a YF virus recovery of only about 20 percent (%); and the final virus product for dosing includes a relatively high amount of Host Cell Protein (HCP).
  • HCP Host Cell Protein
  • a typical YF vaccine dose for a phase 1 clinical trial may be about 0.5 milliliters (ml) of YF virus in a suspension.
  • the remaining HCP in a dose purified by prior art methods may typically be about forty-five thousand nanograms (45,000 ng/dose) wherein the dose is about 8.3 log10 Viral Equivalents (VE).
  • VE is the Elisa unit for YF.
  • the disclosed method provides, for the modified YF virus purification tests run to date, a downstream virus recovery of from about 30 percent to about 100 percent; and for a dose of about 0.5 milliliters (ml) of YF virus, a HCP level that is below the limit of detection of the commercially used vero cell HCP assay.
  • Modified Yellow fever virus was prepared from the attenuated YF 17D virus available commercially as the vaccine, YF-VAX® (Sanofi Pasteur, Swiftwater Pa.).
  • the attenuated YF 17D virus was adapted by serial passage to replicate more efficiently in Vero cells derived from the WHO Vero 10-87 Cell Bank and passaged in serum free medium. 10 serial passages were used to modify the nucleotide sequence of the viral genome virus to develop a seed virus with enhanced growth in Vero cells for preparation of an inactivated Yellow Fever virus candidate.
  • the master and working virus seeds were manufactured from the conditioned cell culture medium harvested from stationary cultures of Vero cells prepared from the Manufacturers' Working Cell bank (MWCB).
  • MWCB Manufacturers' Working Cell bank
  • the working virus seed was used to infect Vero cells prepared from the MWCB grown on CYTODEXTM 1 microcarriers in either a 50 liter, single use bioreactor (working volume 25 to 40 L) or a 10L glass bioreactor (working volume 8 L) (Xcellerex, Inc., Marlborough, Mass.).
  • the virus released into the cell culture medium was harvested from about 5 to about 7 days after infection. From about 2 days to about 3 days before harvest of the virus, the culture was re-fed with fresh medium. This re-feeding step has been shown to increase virus yield. See PCT/US2010/043013, “Drain Down and Re-Feed of Microcarrier Bioreactor,” filed 23 Jul. 2010 and published 27 Jan. 2011 as WO 2011/011660, the entire teachings of which are incorporated herein by
  • the New Downstream Process outlined in the right side of FIG. 6 can have the following steps, each represented by a rectangle in the Figure: Following Virus Harvest, (1) Clarification using a depth filter, and buffer adjustment. (2) BENZONASE® digestion and 0.2 Micron Filtration, producing a live virus bulk. (3) Purification using CELLUFINE® Sulfate Chromatography and Dilution for inactivation step. (4) Virus Inactivation and 0.45 Micron Filtration. (5) Sucrose Gradient UltraCentrifugation. (6) Identify Fractions; Pool Fractions; Warm to from about 25° C. to about 30° C.; 0.2 Micron Filtration, forming Bulk Drug Substance, a purified, inactivated virus. (7) Alum Binding and Formulation.
  • the New and Improved Downstream Process outlined in the right side of FIG. 7 can have the steps as in the New Downstream Process shown in FIG. 6 , with the following changes.
  • GE CAPTOTMCore 7FT with one percent (Human Serum Albumin) HSA or rHSA and adjustment to 20 percent sucrose/0.0005 percent TWEENTM-20.
  • the virus inactivation there is an adjustment with one percent human albumin or rhuman albumin and 0.2 micron filtration to yield the bulk drug substance which can then be formulated with Alum, buffer exchange, and stabilizers.
  • virus data determined by the 2E10 monoclonal antibody is elevated when bound to alum. While not being bound by theory, we postulated that the virus particle is arranged on the surface of the alum hydrogel so that the epitope may be presented in a more open form. Another hypothesis is that the particles are arranged in a more symmetrical manner, thus precluding the formation of aggregates which could mask epitope exposure.
  • the conditioned cell culture medium containing virus was removed from the bioreactor and clarified by depth filtration.
  • a Millipore DE50 depth filter was used to clarify the Virus Harvest.
  • the depth filter was flushed twice, once with USP purified H 2 O followed by buffer with the target formulation 20 mM Tris, 145 mM NaCl, pH 8.
  • the harvest material was passed through the depth filter at a flow rate of approximately 500 mL/min and a pressure not to exceed 25 psi.
  • Filtered material was collected into a bioprocess single-use bag.
  • the depth filter was chased with the same buffer and the chase volume was combined with the original filtrate.
  • the material was adjusted to a target formulation of 50 mM Tris, pH 8 and 2 mM MgCl2 in preparation for the subsequent BENZONASE® treatment step.
  • the adjusted clarified harvest was mixed for approximately 10 min at room temperature.
  • the adjusted clarified virus intermediate was treated with BENZONASE® in order to fragment Vero cell DNA.
  • BENZONASE® was added to the adjusted clarified virus to a final target concentration of 3 units/mL, and the suspension was mixed for 16 to 18 hours at room temperature. After BENZONASE® treatment the product pool was 0.5 ⁇ m filtered.
  • CELLUFINE® sulfate (Chisso, Tokyo, Japan) is a virus affinity resin designed to concentrate, purify and depyrogenate virus. This process step significantly reduces Vero cell proteins, endotoxin and Vero cell DNA.
  • the 2E10 ELISA that detects a YF viral envelope epitope is used to measure the virus concentration on a sample of the clarified and BENZONASE®-treated live virus to ensure the column is appropriately sized to process a virus mass challenge of no more than 5-6E+09 VE per /ml of CELLUFINE® sulfate resin.
  • the column Prior to loading the live virus material, the column was charged with 0.1 M NaoH/0.5 M NaCl buffer at an approximate linear flow rate of 200 cm/hr. The column was then equilibrated with equilibration buffer, 10 mM Tris, 145 mM NaCl, pH 7.5 at an approximate linear flow rate of 200 cm/hr. Post equilibration, the appropriate volume of the virus was loaded onto the column at an approximate linear flow rate of 200 cm/hr.
  • the column was washed with 10 mM Tris, 145 mM NaCl, pH 7.5 buffer at an approximate linear flow rate of 200 cm/hr.
  • the bound virus was eluted from the column using 10 mM Tris, 1.5 M NaCl, pH 7.5 buffer at a reduced linear flow rate of approximately 100 cm/hr. Decreasing the elution flow rate increases buffer residence time and, therefore, decreases elution volume.
  • the elution pool was immediately diluted 2 ⁇ (1 part eluate to 1 part dilution buffer) with 62.5 mM HEPES, pH 8 (target formulation) buffer to reduce precipitation of virus.
  • the resin was cleaned with 0.1 N NaOH/0.5 M NaCl at an approximate linear flow rate of 200 cm/hr.
  • the column was stored in 0.1 N NaOH/0.5 M NaCl at room temperature.
  • the 2 ⁇ diluted CELLUFINE® Sulfate elution pool was then loaded onto a CAPTOTM Core 700 column. Prior to loading, the column was regenerated with 0.1 N NaOH/0.5M NaCl at a linear flow rate of 300 cm/hr.
  • One column volume of a re-equilibration buffer of 500 mM Tris/145 mM NaCl, pH 7.5 was applied prior to the equilibration buffer consisting of 20 mM MES/100 mM NaCl, pH 7 at a linear flow rate of 300 cm/hr.
  • the column was washed with 20 mM MES/100 mM NaCl, pH 7 at a linear flow rate of 300 cm/hr. The flow through and wash was collected as the product. This fraction was then diluted 2 ⁇ with 50 mM HEPES/20% Sucrose/0.001% TWEENTM-20, pH 8. The resin was cleaned with 1 N NaOH/1M NaCl at a linear flow rate of 300 cm/hr. The column was stored in 0.1 N NaOH/0.5M NaCl at room temperature.
  • a 10% solution of BPL was made by diluting BPL with water for injection (WFI).
  • WFI water for injection
  • the 10% BPL was stored in single use aliquots at ⁇ 60° C.
  • the concentration of BPL in this 10% solution was confirmed by gas chromatography (GC) analysis.
  • HSA Human serum albumin
  • the inactivation mixture was mixed for approximately 3 hours at room temperature on a low heat-generating stir plate. This material was then incubated at 30° C. for 60 minutes, and filtered using a 0.2 ⁇ m PES filter.
  • the 0.2 ⁇ m filtered purified inactivated virus was bound to “alum” [Aluminum aluminum Hydroxide hydroxide (Alum ALHYDROGEL®)] and buffer exchanged into the final formulation buffer. All process steps were aseptically performed.
  • Alum alum was added to 9 parts of 0.2 ⁇ m filtered sucrose gradient-purified inactivated virus to achieve a final alum target concentration of 0.2%.
  • the resulting product was mixed for a target of 1-4 hours at room temperature.
  • the alum-bound virus was aseptically buffer exchanged into 10 mM Tris/1.2 mM MgCl2/10 mM L-glutamic acid/0.11 mM D-mannitol/2 mM Trimethylamine-N-oxide dihydrate, pH 7.5.
  • the alum-bound virus was settled by centrifugation or membrane filtration. After settling, the supernatant was decanted.
  • the alum-bound virus pellet was re-suspended with formulation buffer to a volume equal to the original alum-bound virus pool volume and mixed for a target of 10 minutes at room temperature. This process was repeated 3-4 ⁇ until the alum-bound virus pool was exchanged into the final formulation buffer.
  • the alum-bound and buffer exchanged virus pool was stored at 2-8° C. This is referred to herein as the “Bulk Drug Product.”
  • the potency of the Bulk Drug Product was measured using ELISA that detects alum-bound YF.
  • a process recovery chromatogram for Experiment 1639A using a CELLUFINE® Sulfate Column is shown in FIG. 4 and for Experiment 1639A using a CAPTOTM DeVirS column in FIG. 5 .
  • FIG. 8 Two other embodiments of the disclosed process are shown in FIG. 8 , “Downstream new and Improved process #3.”
  • the disclosed chromatographic purification process yielded sufficient virus recovery and reduced the host cell protein levels to below the limit of detection of the commercial Vero cell HCP assay. These virus recoveries and HCP results were comparable to results achieved by a similar purification process utilizing sucrose gradient ultracentrifugation.
  • a benefit of the disclosed process is that, unlike centrifugation methods, it is suitable for use in single-use systems.
  • the next phase of the Yellow Fever purification development is to replace the Chisso CELLUFINE® Sulfate resin with an equivalent GE Healthcare resin.
  • the CELLUFINE® sulfate resin can potentially be replaced with GE Healthcare's CAPTOTM DeVirS resin.
  • the CAPTOTM DeVirS is part of GE Healthcare's Custom Designed Media program, and is an affinity chromatography resin with the ligand dextran sulfate, which is known to have an affinity-like behavior to different types of virus.
  • CAPTOTM DeVirS offers the a number of benefits for purification of virus, including, for example, excellent productivity, good chemical stability, and affinity-like behavior to various viruses.
  • next planned experiments include following the use of CAPTOTM DeVirS with GE CAPTOTM Core 700 .
  • anion exchange membrane such as a Q Monolith (BIA) or a Q membrane (Natrix, Pall, Sartorius) between the step using the CAPTOTM DeVirS and the step using GE CAPTOTM Core 700.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Virology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
US14/423,394 2012-08-24 2013-08-16 Virus purification and formulation process Abandoned US20150210986A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US14/423,394 US20150210986A1 (en) 2012-08-24 2013-08-16 Virus purification and formulation process

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201261692956P 2012-08-24 2012-08-24
PCT/US2013/055302 WO2014031480A1 (en) 2012-08-24 2013-08-16 Virus purification and formulation process
US14/423,394 US20150210986A1 (en) 2012-08-24 2013-08-16 Virus purification and formulation process

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2013/055302 A-371-Of-International WO2014031480A1 (en) 2012-08-24 2013-08-16 Virus purification and formulation process

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US15/163,288 Continuation US10196615B2 (en) 2012-08-24 2016-05-24 Virus purification and formulation process

Publications (1)

Publication Number Publication Date
US20150210986A1 true US20150210986A1 (en) 2015-07-30

Family

ID=48998541

Family Applications (4)

Application Number Title Priority Date Filing Date
US14/423,394 Abandoned US20150210986A1 (en) 2012-08-24 2013-08-16 Virus purification and formulation process
US15/163,288 Active US10196615B2 (en) 2012-08-24 2016-05-24 Virus purification and formulation process
US16/229,836 Abandoned US20190136206A1 (en) 2012-08-24 2018-12-21 Virus purification and formulation process
US18/085,110 Pending US20230122337A1 (en) 2012-08-24 2022-12-20 Virus purification and formulation process

Family Applications After (3)

Application Number Title Priority Date Filing Date
US15/163,288 Active US10196615B2 (en) 2012-08-24 2016-05-24 Virus purification and formulation process
US16/229,836 Abandoned US20190136206A1 (en) 2012-08-24 2018-12-21 Virus purification and formulation process
US18/085,110 Pending US20230122337A1 (en) 2012-08-24 2022-12-20 Virus purification and formulation process

Country Status (6)

Country Link
US (4) US20150210986A1 (pt)
EP (1) EP2700711B1 (pt)
AU (1) AU2013306080B2 (pt)
BR (1) BR102013021499B1 (pt)
WO (1) WO2014031480A1 (pt)
ZA (1) ZA201501601B (pt)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113166732A (zh) * 2018-12-20 2021-07-23 怡诺安有限公司 使用亲和色谱法的用于疫苗病毒的纯化方法
US11162880B2 (en) 2015-11-09 2021-11-02 University Of Notre Dame Du Lac Particle size purification method and devices

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014031480A1 (en) * 2012-08-24 2014-02-27 Xcellerex, Inc. Virus purification and formulation process
CA2980812A1 (en) * 2015-04-03 2016-10-06 Robert Schlegl Aseptic purification process for viruses

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1724338A1 (en) * 2005-05-19 2006-11-22 Crucell Holland B.V. Methods for the production of a whole-inactivated West Nile Virus vaccine

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10144903A1 (de) * 2001-09-12 2003-03-27 Chiron Behring Gmbh & Co Vermehrung von Viren in Zellkultur
JP4629660B2 (ja) * 2003-03-05 2011-02-09 ジーイー・ヘルスケア・バイオサイエンス・アクチボラグ マルチモードアニオン交換リガンドの製造法
PL2007883T3 (pl) * 2006-04-20 2012-07-31 Wyeth Llc Sposób oczyszczania do izolacji oczyszczonego wirusa pęcherzykowatego zapalenia jamy ustnej z hodowli komórkowej
WO2011011660A2 (en) 2009-07-23 2011-01-27 Xcellerex, Inc. Drain down and re-feed of microcarrier bioreactor
CA3150411C (en) * 2009-07-31 2023-09-12 Pnuvax Inc. High yield yellow fever virus strain with increased propagation in cells
AR079970A1 (es) * 2010-07-23 2012-02-29 Xcellerex Inc Cepas de alto rendimiento del virus de la fiebre amarilla con una propagacion incrementada en las celulas
WO2014031480A1 (en) * 2012-08-24 2014-02-27 Xcellerex, Inc. Virus purification and formulation process

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1724338A1 (en) * 2005-05-19 2006-11-22 Crucell Holland B.V. Methods for the production of a whole-inactivated West Nile Virus vaccine

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11162880B2 (en) 2015-11-09 2021-11-02 University Of Notre Dame Du Lac Particle size purification method and devices
CN113166732A (zh) * 2018-12-20 2021-07-23 怡诺安有限公司 使用亲和色谱法的用于疫苗病毒的纯化方法
TWI803725B (zh) * 2018-12-20 2023-06-01 南韓商怡諾安有限公司 使用親和層析法之用於疫苗病毒的純化方法

Also Published As

Publication number Publication date
US20160264943A1 (en) 2016-09-15
US20190136206A1 (en) 2019-05-09
BR102013021499B1 (pt) 2022-02-08
WO2014031480A1 (en) 2014-02-27
AU2013306080B2 (en) 2019-01-31
US20230122337A1 (en) 2023-04-20
US10196615B2 (en) 2019-02-05
AU2013306080A1 (en) 2015-02-26
BR102013021499A2 (pt) 2016-09-20
ZA201501601B (en) 2016-01-27
EP2700711A1 (en) 2014-02-26
EP2700711B1 (en) 2017-11-22

Similar Documents

Publication Publication Date Title
US20230122337A1 (en) Virus purification and formulation process
JP5129805B2 (ja) 細胞培養物から精製水疱性口内炎ウイルスを単離するための精製プロセス
CN106574252B (zh) 从细胞培养物中纯化脊髓灰质炎病毒的方法
RU2565827C2 (ru) Способ получения вирусного антигена и вакцин
Trabelsi et al. Purification of rabies virus produced in Vero cells grown in serum free medium
KR20170138462A (ko) 바이러스에 대한 무균 정제 방법
KR19990036028A (ko) 일본뇌염 백신의 공업적인 생산방법 및 이에 의한 백신
CA2859916A1 (en) Purification of herpes virus
EP3121268B1 (en) Methods for purification of a virus produced in vitro and clearance assay for the virus
EP3818150A1 (en) Method for purifying an enveloped virus
US10421986B2 (en) Method for the clarification of high-density crude cell culture harvest
CN110204613B (zh) 一种针对胡宁病毒的中和抗体、其制备方法及其应用
Wang et al. Efficient purification of cell culture-derived classical swine fever virus by ultrafiltration and size-exclusion chromatography
TW202031893A (zh) Vlp表現cho細胞株之建構
CA2951735C (en) Preparation methods for a novel generation of biological safe klh products used for cancer treatment, for the development of conjugated therapeutic vaccines and as challenging agents
CN111979201B (zh) 提高口蹄疫病毒表达水平的方法
Ferreira Overcome challenges in influenza virus-like particles downstream process

Legal Events

Date Code Title Description
AS Assignment

Owner name: XCELLEREX INC., MASSACHUSETTS

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:VICALVI, JOHN;HAYMAN, EDWARD;MAKOWIECKI, JOSEPH;SIGNING DATES FROM 20130816 TO 20150311;REEL/FRAME:038706/0325

AS Assignment

Owner name: GE HEALTHCARE BIO-SCIENCES CORP., MASSACHUSETTS

Free format text: MERGER;ASSIGNOR:XCELLEREX, INC.;REEL/FRAME:038731/0200

Effective date: 20131014

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION