Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
  • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
  • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
  • G01N21/65—Raman scattering
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
  • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
  • G01J3/28—Investigating the spectrum
  • G01J3/44—Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
  • G01J3/4412—Scattering spectrometry
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
  • G01N33/56983—Viruses
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2201/00—Features of devices classified in G01N21/00
  • G01N2201/12—Circuits of general importance; Signal processing
  • G01N2201/126—Microprocessor processing
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2201/00—Features of devices classified in G01N21/00
  • G01N2201/12—Circuits of general importance; Signal processing
  • G01N2201/129—Using chemometrical methods
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2201/00—Features of devices classified in G01N21/00
  • G01N2201/12—Circuits of general importance; Signal processing
  • G01N2201/129—Using chemometrical methods
  • G01N2201/1296—Using chemometrical methods using neural networks

Definitions

• One advantage of the present invention is that viral nucleic acid abundance and viral structural molecule abundance, and the ratio of viral nucleic acids to viruses comprising one or more viral structural molecules, can be continuously monitored in real-time. There is no need to process samples from the viral culture medium to generate an estimate of viral nucleic acid abundance, viral structural molecule abundance and the ratio of viral nucleic acids to viruses comprising one or more viral structural molecules. Measurements may be made in situ , if desirable. In other words, measurements may be made directly on the viral culture medium in the growth incubator. Measurements may be made ex situ , if desirable. In other words, measurements may be made directly on the viral culture medium in an aliquot of the viral culture medium taken from the growth incubator or separated from the main chamber of the growth incubator.
• the change in the ratio in the sample may be used to determine the start phase, the production phase and/or the stationary phase of a viral production process. Any such method may be used to determine the optimal conditions for a viral production process. Any such method may be used to assess a process downstream of a viral production process.
• viral vectors may be based on wild type viruses, they are generally modified as compared to wild type viruses and are commonly used to introduce genetic material into target cells (e.g. genes of therapeutic use). Viral vectors therefore have particular utility, e.g. for gene therapy, cell therapy or for other molecular applications, and their production is of enormous importance to the gene therapy and cell therapy industries. It will be well understood that for example, modifications may be made to improve safety of viral vectors for gene and/or cell therapy or to improve for example the size of gene which may be carried by the vector.
• RNA viral genome and is marked by the presence of the psi sequence which ensures that the genome is subsequently packaged into the virion.
• lentiviral vectors may be produced by the transformation and expression of three (for second generation systems) or four (for third generation systems) plasmids in a producer cell line. Plasmids for the production of viral vectors are commercially available, e.g. Lenti-Pac and AAV Prime (GeneCopoeia).
• “Viral components” are considered herein to be any part of the virus, virus particle or viral vector.
• the Stokes scattered light is monitored as the measured signals are more intense at ambient temperatures.
• Figure 2 shows some example spectra; the different peaks represent the presence of different modes of vibration; some bands are overlapped regions of several underlying peaks.
• the different peaks represent the presence of different modes of vibration; some bands are overlapped regions of several underlying peaks.
• the plurality of wavenumber ranges in the Raman spectrum which are measured may comprise 5 or more of the wavenumber ranges 1 to 28 as listed in Table 3 below and wherein the VIP is > 1.00.
• the plurality of wavenumber ranges in the Raman spectrum which are measured may comprise 10 or more of the wavenumber ranges 1 to 28 as listed in Table 3 below and wherein the VIP is > 1.00.
• the plurality of wavenumber ranges in the Raman spectrum which are measured may comprise 15 or more of the wavenumber ranges 1 to 28 as listed in Table 3 below and wherein the VIP is > 1.00.
• the results show that the model using the Raman spectroscopy data is consistent with offline measurements of viral titre over time.
• a comparison of the titres obtained from the RT-qPCR assay and P24 ELISA are shown in Figure 10.
• AAV8 particles were detected using two steps 1) a biotin- conjugated anti-AAV8 antibody was bound to the immune complex 2) a streptavidin peroxidase conjugate reacts with the biotin molecules. Addition of the tetramethylbenzidine (TMB) substrate solution resulted in a colour reaction, which is proportional to the amount of specifically bound viral particles. The absorbance is then measured photometrically at 450nm.
• TMB tetramethylbenzidine

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• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Physics & Mathematics (AREA)
• Immunology (AREA)
• Chemical & Material Sciences (AREA)
• General Physics & Mathematics (AREA)
• Engineering & Computer Science (AREA)
• Biochemistry (AREA)
• Analytical Chemistry (AREA)
• General Health & Medical Sciences (AREA)
• Pathology (AREA)
• Urology & Nephrology (AREA)
• Molecular Biology (AREA)
• Hematology (AREA)
• Biomedical Technology (AREA)
• Spectroscopy & Molecular Physics (AREA)
• Virology (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Biotechnology (AREA)
• Medicinal Chemistry (AREA)
• Food Science & Technology (AREA)
• Microbiology (AREA)
• Cell Biology (AREA)
• Tropical Medicine & Parasitology (AREA)
• Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
• Investigating Or Analysing Biological Materials (AREA)
• Apparatus Associated With Microorganisms And Enzymes (AREA)
• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

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Publications (1)

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Cited By (5)

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• 2020
  • 2020-07-01 GB GBGB2010104.4A patent/GB202010104D0/en not_active Ceased
• 2021
  • 2021-07-01 JP JP2022581700A patent/JP2023532568A/ja active Pending
  • 2021-07-01 AU AU2021301333A patent/AU2021301333A1/en not_active Abandoned
  • 2021-07-01 IL IL299065A patent/IL299065A/en unknown
  • 2021-07-01 WO PCT/GB2021/051673 patent/WO2022003359A1/en not_active Ceased
  • 2021-07-01 KR KR1020237003599A patent/KR20230028556A/ko active Pending
  • 2021-07-01 EP EP21742868.9A patent/EP4176245A1/en active Pending
  • 2021-07-01 CA CA3182045A patent/CA3182045A1/en active Pending
  • 2021-07-01 US US18/003,629 patent/US20230236128A1/en active Pending
  • 2021-07-01 CN CN202180052867.XA patent/CN116034263A/zh active Pending

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