2021 Gene and Cell Therapy Calendar - Vigene Biosciences
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2021 Gene and Cell Therapy Calendar
V
igene Biosciences, an award-winning world leader in plasmid and
viral vector development and manufacturing, wishes you a healthy
and prosperous 2021.
This 2021 calendar contains figures and tables that have been
carefully selected by Vigene from gene and cell therapy articles, all reprinted with
permission from Springer Nature.
We hope that it will be a useful reference for researchers in the gene and cell
therapy field. Vigene Bioscience has been ranked by Inc. Magazine as one of
the fastest growing companies in the USA for the past three years. Vigene offers
integrated plasmid and viral vector production and analytical services from
its 71,000 sq ft state-of-the-art facility which includes 10 Good Manufacturing
Practice (GMP) clean-room suites. Vigene’s mission is to make gene therapy
affordable. On the basic research side, Vigene is developing, manufacturing, and
distributing adeno-associated virus (AAV), lentivirus, retrovirus, adenovirus, and
plasmid-based reagents including Howard Hughes Medical Institute (HHMI)/
Janelia Research Campus AAV Biosensors. On the cGMP clinical production
side, Vigene combines proven production technologies with rigorous, regulatory
compliant cGMP production processes to meet the needs and expectations of
clinical and commercial clients.
Vigene offers FDA and EMA compliant cGMP production for AAV,
lentivirus, adenovirus, retrovirus, and plasmids to global pharmaceutical and
biotech companies, government agencies, and non-profit organizations.Viral Vector Genome Size Infec on Expression Pote a ons
onal mutagenesis
Retrovirus 7-11 kb (ssRNA) Dividing cells Stable
poten al
Dividing & non- onal mutagenesis
Len virus 9 kb (ssRNA) Stable
dividing cells poten al
Dividing & non-
Adenovirus 36 kb (dsDNA) Transient Immune response
dividing cells
Adeno-
Dividing & non-
associated 4.7 kb (ssDNA) long las ng Immune response
dividing cells
Virus, AAV
Herpes Simplex Dividing & non- No gene expression
150 kb (dsDNA) Transient
Virus, HSV dividing cells during latent infec on
Pote al cytopathic
Vaccinia virus 190 kb (dsDNA) Dividing cells Transient
effects
Vigene integrated vector and plasmid service — Research, preclinical and clinical
Vigene is a world leader in plasmid and viral vector development and manufacturing. Vigene has expertise
in development, manufacturing and analytics for plasmid vectors and many viral vectors, including AAV,
lentivirus, adenovirus, retrovirus, HSV, and vaccinia virus.
Please see the Jan 2021 promotion here: vigenebio.com/2021/JanVigene integrated vector and plasmid service
MONDAY TUESDAY WEDNESDAY THURSDAY
JANUARY
FRIDAY
2021
SATURDAY SUNDAY
1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31
www.vigenebio.comTransmission electron microscopy of rAAV. It is clear that full particles (open arrow)
stain differently from empty particles (darkly stained center; small dark arrow).
Viewing numerous fields similar to this will allow determination of the full-to-empty
particle ratio. (Figure 8, Nature Protocols 1, 1412–1428 (2006))
Figure reprinted with permission from Springer Nature
AAV reference standards
The gene and cell therapy field needs good AAV reference standards. Pursuant to the
Jan 2020 FDA Gene Therapy Chemistry Manufacturing and Controls (CMC) guidance,
Vigene’s AAV reference standards are as follows:
• AAV1, AAV2, AAV5, AAV6, AAV8, and AAV9 serotypes
• Full and empty capsids
• AAV titer standards
Please view the details and promotions at: vigenebio.com/2021/FebAAV reference standards
MONDAY TUESDAY WEDNESDAY
FEBRUARY
THURSDAY FRIDAY
2021
SATURDAY SUNDAY
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
www.vigenebio.comITR ITR AAV vector packaging
(4.5-kb capacity
without engineering)
Promoter Therapeutic transgene Poly(A)
5′ 3′ OH
Recombinant
AAV virion
• Screen for and Use of dual vectors that take
• Mutation of ITRs • Codon optimization
Engineering use small, strong, advantage of:
to generate self- via codon usage bias • Synthetic poly(A)
approach tissue-specific • AAV genome concatemerization
complementary • Interference of • Reversed poly(A)
promoters and • Homologous recombination
AAV intermediates antigen presentation
enhancer elements • A hybrid dual-vector strategy
• Intein-mediated protein
• Increases • Enhances trans-splicing technology
tissue-specific polyadenylation Cross-packaging of an AAV
• Omits need transcription • Increases • Minimizes kilobases of genome
Positive
for second- of transgene transcription DNA taken up by poly(A)
effect on
strand synthesis • Minimizes and translation of • Avoids rolling circle
transduction
of ssDNA kilobases of DNA the transgene transcription and Pol II
taken up by jumping (between • Increases size of transgene that
promoter concatamers) can be packaged
• Decreases innate
Positive
• Decreases immune response • Decreases innate
effect on • Decreases CTL
innate immune to AAV immune response
immune response to AAV
response to AAV • Reduces CTL to AAV
response
response
Engineering the AAV cassette. The adeno-associated virus (AAV) cassette can be engineered to enhance AAV transduction and also to enable AAV to
escape immune responses. Mutation of one inverted terminal repeat (ITR) on the AAV vector, which prevents the nicking of Rep protein, can generate
self-complementary AAV vector to enhance vector transduction. Mutation of ITRs may also decrease the innate response to AAV. Use of small
tissue-specific promoters increases tissue-specific transgene expression and the packaging capacity of the AAV genome and minimizes the cytotoxic
T lymphocyte (CTL) immune response to AAV. Optimization of transgene codons increases the transcription and translation of the AAV transgene
and decreases the immune response to AAV. Using synthetic poly(A) can increase the nuclear export, translation and stability of mRNA (by enhancing
polyadenylation), and using reversed poly(A) can avoid the transcription of ITR; both approaches enhance AAV transduction, and reversed poly(A)
decreases the innate response to AAV. Finally, the use of dual AAV vectors or the cross-packaging of the AAV genome enables effective and functional
expression of large transgenes. Pol II, DNA polymerase II; ssDNA, single-stranded DNA. (Figure 2, Nature Reviews Genetics 21, 255–272 (2020))
Figure reprinted with permission from Springer Nature
Plasmid production – Research, preclinical, clinical
• Research grade & GMP-ReadyTM, GMP production
• Ready-to-use, GMP-Ready pHelper & other viral vector packaging plasmids
• Endotoxin free & supercoil plasmid homogeneity
• Animal component free production process
Please view the details at vigenebio.com/2021/MarPlasmid production – preclinical and GMP
MONDAY TUESDAY WEDNESDAY THURSDAY
MARCH
FRIDAY
2021
SATURDAY SUNDAY
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30 31
www.vigenebio.comTABLE 1: AAV PRODUCTION OPTIONS
Pros and cons of various recombinant adeno-associated virus (rAAV) manufacturing strategies. rAAV cannot replicate
without a helper virus, and early manufacturing efforts entailed coinfection of host cells with adenovirus or herpesvirus. Newer
strategies replace these with plasmids containing key helper virus genes, or combine all necessary genetic elements into an
insect cell-specific baculovirus vector. Production is most efficient in free-floating suspension cells, but substrate-attached
adherent cell lines can also achieve reasonable viral output.
AAV MANUFACTURING KEY KEY PRODUCTION CELL LINE CHOICES
TECHNOLOGY STRENGTHS DRAWBACKS
ADHERENT SUSPENSION
• Helper virus
• Highly scalable contamination
• Serum-free media • Long lead time for cell HEK293/293T HEK293/293T-s
Helper virus • Efficient line and virus seed
production in generation HeLa HeLa-s
suspension culture • May require serum-
containing media
• May require serum-
• No helper virus containing media
contamination • Large proportion of
Helper-free triple
• Rapidly produce empty capsids HEK293/293T HEK293/293T-s
transfection
virus in small scale • Supply of plasmids for
• Simple procedure large-scale production
can be costly
• Baculovirus virus
• Highly scalable contamination
Serum-free media
• Baculovirus instability
Baculovirus • Efficient — sf9
production in • Long lead time for cell
suspension culture line and virus seed
generation
AAV production – Research, preclinical, clinical
• Research grade, preclinical, clinical, and commercial GMP AAV production
• Proprietary cGMP released MCB of HEK293, 293T adherent and suspension cell lines
• Established and proven high productivity AAV production process
• >15,000 research batches, >50 preclinical and clinical batches released
Please view the promo at vigenebio.com/2021/AprAAV production – preclinical and GMP
MONDAY TUESDAY WEDNESDAY THURSDAY
APRIL
FRIDAY
2021
SATURDAY SUNDAY
1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30
www.vigenebio.comTarget cell
Virus-mediated transduction is a
useful tool to generate various
types of next-generation stem cells
Virus as a key tool
Click chemistry
Nuclease or AAV-based gene editing
a Prodrug-converting enzymes b Optogenetic actuator-embedded c CAR-expressing HSCs or
or oncolytic virus delivery via stem cell derivative PSC-derived NK cells
tumour-homing NSCs/MSCs CAR
Stem cell
Oncolytic virus
particles (or prodrug-
converting enzymes)
e Cytokine/growth factor-overexpressing
stem cells to enhance endogenous repair
d Gene therapy of stem cells to provide
long-term functional compensation
for disease-causing, inherited mutation
f Improved delivery of anticancer g Gene editing tools can be used to knock
agents to sites of action using click out expression of HLA genes to reduce the
chemistry to tether them to stem cells immunogenicity of allogeneic stem cells
Azide
DBCO
HSC
Platelet
Anti-PD-L1
The stem cell toolkit and its application in developing next-generation stem cells. Virus-mediated transduction of stem cells (target cell), particularly with self-inactivating lentiviruses,
is a useful tool for the creation/development of next-generation stem cells. Viruses can be used to engineer: prodrug-converting enzymes, oncolytic viruses and other anticancer drugs
into neural stem cells (NSCs) and mesenchymal stem cells (MSCs) (part a); optogenetically enhanced stem cell derivatives to provide light-inducible control over the activity of trans-
planted stem cells/progenitors (part b); chimeric antigen receptor (CAR)-expressing haematopoietic stem cells (HSCs) and pluripotent stem cell (PSC)-derived natural killer (NK) cells
for immune-oncology applications (part c); gene therapy in HSC, skin and muscle progenitors to treat inherited diseases (part d); and cytokine/growth factor delivery in MSCs/neural
progenitors to stimulate endogenous tissue repair (part e). Click chemistry is another engineering tool that can be used to tether anticancer agents to stem cells for improved delivery to
hard-to-reach cancers, such as leukaemia cells residing deep in the bone marrow (part f). Advances in gene editing technology have made this a versatile tool to precisely edit specific
loci within the genome, and gene editing technology has become the tool of choice for the creation/development of universally immunocompatible PSC lines and derivatives (part g).
Several of these next-generation stem cell-based therapies have already reached clinical testing, whereas others in preclinical development are not far behind. AAV, adeno-associated
virus; DBCO, dibenzocyclooctyne; HLA, human leukocyte antigen; PD-L1, programmed cell death protein 1 ligand. (Figure 1, Nature Reviews Drug Discovery 19, 463–479 (2020))
Figure reprinted with permission from Springer Nature
Lentivirus production – Research, preclinical, clinical
• Research grade, preclinical, clinical, and commercial GMP lentivirus production
• High titer, purified, ready to use in vitro and animal studies — research grade
• Chromatography based commercial ready lenti production — GMP
• High transduction efficiency
Please view the promo at vigenebio.com/2021/MayLentivirus research production
MONDAY TUESDAY WEDNESDAY THURSDAY
MAY
FRIDAY
2021
SATURDAY SUNDAY
1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
www.vigenebio.comOncolytic Cytokine
virus receptors
Release/
secrete
Infection Cancer cell NK cell
Cytotoxicity
• Viral proteins
• Viral genome
CD8+ T cell
• ER stress
ROS Cytokine
• Genotoxic stress
receptors IL-2R
Cytotoxicity
IL-2
Viral
oncolysis ROS
Release Release CD28 TCR
CD4+ T cell
Antigen
uptake
MHC MHC CD40L
TLR
PAMPs DAMPs Cytokines
• Viral capsids • HSPs • Type I interferons
• Viral DNA • HMGB1 • TNFα Activation CD40
• Viral dsRNA/ssRNA • Calreticulin • IFNγ TCR
• Viral proteins • ATP • IL-12
• Uric acid
MHC
• Type I IFNs • Type I IFNs DAMPs/
• DAMPs/PAMPs • Cytokines PAMPs
• Viral antigens • CD80/CD86
• TAAs/neoantigens • Chemokine receptors
Antigen presenting cell
The induction of local and systemic anti-tumour immunity by oncolytic viruses. The therapeutic efficacy of oncolytic viruses is determined by a combination of direct cancer
cell lysis and indirect activation of anti-tumour immune responses. Upon infection with an oncolytic virus, cancer cells initiate an antiviral response that consists of endoplasmic
reticulum (ER) and genotoxic stress. This response leads to the upregulation of reactive oxygen species (ROS) and the initiation of antiviral cytokine production. ROS and
cytokines, specifically type I interferons (IFNs), are released from the infected cancer cell and stimulate immune cells (antigen presenting cells, CD8+ T cells, and natural killer
(NK) cells). Subsequently, the oncolytic virus causes oncolysis, which releases viral progeny, pathogen-associated molecular patterns (PAMPs), danger-associated molecular
pattern signals (DAMPs), and tumour associated antigens (TAAs) including neo-antigens. The release of viral progeny propagates the infection with the oncolytic virus. The
PAMPs (consisting of viral particles) and DAMPs (comprising host cell proteins) stimulate the immune system by triggering activating receptors such as Toll-like receptors
(TLRs). In the context of the resulting immune-stimulatory environment, TAAs and neo-antigens are released and taken up by antigen presenting cells. Collectively, these events
result in the generation of immune responses against virally infected cancer cells, as well as de novo immune responses against TAAs/neo-antigens displayed on un-infected
cancer cells. CD40L, CD40 ligand; dsRNA, double-stranded RNA; HMGB1, high mobility group box 1; HSP, heat shock protein; IL-2, interleukin-2; IL-2R, IL-2 receptor; MHC, major
histocompatibility complex; ssRNA, single-stranded RNA; TCR, T cell receptor; TNFα, tumour necrosis factor-α. (Figure 2, Nature Reviews Drug Discovery 14, 642–662 (2015))
Figure reprinted with permission from Springer Nature
Adenovirus production - Research, preclinical, clinical
• Research grade, preclinical, clinical, and commercial GMP adenovirus production
• High titer, purified, ready to use in vitro and animal studies – research grade
• Chromatography based commercial ready adenovirus production – GMP
• Oncolytic virus and vaccine production process ready
Please view the promo at vigenebio.com/2021/JuneAdenovirus production
MONDAY TUESDAY WEDNESDAY THURSDAY
JUNE
FRIDAY
2021
SATURDAY SUNDAY
1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
www.vigenebio.comTable 1 | Properties
Properties and clinical
anduse of AAV
clinical use serotypes
of AAV serotypes
AAV Origin of Primary Co-receptor Tissue tropism Condition Approved drug
serotype isolation receptor (ClinicalTrials.gov identifier)
AAV1 Monkey Sialic acid AAVR Muscle, CNS, heart Muscle diseases (NCT01519349) None
Heart failure (NCT01643330)
AAT deficiency (NCT01054339,
NCT00430768)
AAV2 Human Heparin Integrin, FGFR , HGFR, Liver, CNS, muscle Eye diseases (NCT00643747) Luxturna for Leber
LamR, AAVR congenital amaurosis
Haemophilia (NCT00515710)
CNS diseases (NCT00400634)
AAT deficiency (NCT00377416)
AAV3 Human Heparin FGFR , HGFR LamR, Muscle, stem cells No trials underway None
AAVR
AAV4 Monkey Sialic acid Unknown Eye, CNS Eye diseases (NCT01496040) None
AAV5 Human Sialic acid PDGFR, AAVR CNS, lung, eye Haemophilia (NCT03520712) None
Eye diseases (NCT02781480)
AIP (NCT02082860)
AAV6 Human Heparin, EGFR , AAVR Muscle, CNS, heart, Haemophilia (NCT03061201) None
sialic acid lung
CNS diseases (NCT02702115)
AAV7 Monkey Unknown Unknown Muscle, CNS No trials underway None
AAV8 Monkey Unknown LamR, AAVR Liver, muscle, Eye diseases (NCT03066258) None
pancreas, CNS
Haemophilia (NCT00979238)
Muscle diseases (NCT03199469)
AAV9 Human Galactose LamR, AAVR Every tissue CNS diseases (NCT02122952) Zolgensma for spinal
muscular atrophy
Muscle diseases (NCT03362502)
AAV10 Monkey Unknown Unknown Muscle No trials underway None
AAV11 Monkey Unknown Unknown Unknown No trials underway None
AAV12 Human Unknown Unknown Nasal No trials underway None
AAT,α1-antitrypsin; AAV, adeno-associated virus; AAVR, AAV receptor; AIP, acute intermittent porphyria; CNS, central nervous system; EGFR, epidermal growth factor
AAT, α1-antitrypsin; AAV, adeno-associated virus; AAVR, AAV receptor; AIP, acute intermittent porphyria; CNS, central nervous system; EGFR,
receptor ; FGFR , fibroblast growth factor receptor ; HGFR, hepatocyte growth factor receptor ; LamR, laminin receptor 1; PDGFR , platelet-derived growth factor receptor.
epidermal growth factor receptor; FGFR, fibroblast growth factor receptor; HGFR, hepatocyte growth factor receptor; LamR, laminin receptor 1;
PDGFR, platelet-derived growth factor receptor. (Table 1, Nature Reviews Genetics 21, 255–272 (2020))
Table reprinted with permission from Springer Nature
AAV, lentivirus and adenovirus controls
• Reporters available: RFP, GFP, mCherry, Luciferase, Cre, LacZ
• Promoters available: ALB, aMHC, c-Fos, CAG, CaMKIIa, CK0.4, CK1.3, CMV, cTnT, EF1a, EFFS,
GFAP, HCRApoE, MBP, MCK, MeCP2, NSE, PDX1, PGK, Rpe65, SST, Syn, TBG, UBC, DIO-GFP,
DIO-mCherry, DIO-RFP, DIO-LacZ
Please view the promo at vigenebio.com/2021/JulyAAV controls and lentivirus control
MONDAY TUESDAY WEDNESDAY THURSDAY FRIDAY
JULY 2021
SATURDAY SUNDAY
1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31
www.vigenebio.comTable 1 Example
Example of characterization testing
of characterization for for
testing an an
HEK293
HEK293master
master cell bank
cell bank
Test M et hod S pecifi cat ion
Microbial
Fo r m icr o b ia l co n t a m in a t io n Tr a n s m is s io n e le ct r o n m icr o s co p y No viruses, virus-like particles, mycoplasmas,
fungi, yeasts, bacteria
Bacteriostatic/fungistatic activity of test article Four media, direct inoculation No bacterial and fungal activity
Bacterial and fungal contaminants Four media, direct inoculation Negative
Agar cultivable and noncultivable mycoplasmas 1993 points to consider Negative
General viruses
Inapparent viruses In vivo Not detected
Viral contaminants In vitro assay for the presence of Not detected
viral contaminants
Specific human viruses
CM V PCR N e g a t iv e
EBV PCR N e g a t iv e
H AV RT-P C R N e g a t iv e
H BV PCR N e g a t iv e
H CV RT-P C R N e g a t iv e
H H V-6 PCR N e g a t iv e
H H V-7 PCR N e g a t iv e
H H V-8 PCR N e g a t iv e
H IV-1/ 2 PCR N e g a t iv e
H u m a n p a r v o v ir u s B19 PCR N e g a t iv e
H TLV I/ II PCR N e g a t iv e
Re t r o v ir u s e s Q -P ERT < 5.0 10 7
U ml 1
Specific simian viruses
SFV PCR N e g a t iv e
SRV RT-P C R N e g a t iv e
STLV PCR N e g a t iv e
SV40 PCR N e g a t iv e
Other specific viruses
Bovine viruses In vitro assay for the presence of Not detected
bovine viruses (9CFR)
Porcine viruses In vitro assay for the presence of Not detected
porcine viruses (9CFR)
Identity
C e ll id e n t ifi ca t io n Is o e n z y m e a n a ly s is Is o e n z y m e m i g r a t i o n d i s t a n c e s c o n s i s t e n t
with cells of human origin
Abbreviations: CFR, Code
Abbreviations: CFR, of Federal
Code of Regulations;CMV,
Federal Regulations; CMV,cytomegalovirus;
cytomegalovirus; EBV,
EBV, Epstein–Barr
Epstein–Barr virus;
virus; HAV,HAV, hepatitis
hepatitis A virus;
A virus; HBV,HBV, hepatitis
hepatitis B B virus;
HCV,
virus;hepatitis C virus;
HCV, hepatitis HHV,HHV,
C virus; human herpes
human virus;
herpes HIV-1/2,
virus; human
HIV-1/2, immunodeficiency
human immunodeficiency virus
virus types
types 11 and 2; HTLV
and 2; HTLV I/II,
I/II, human T-cell lymphotropic
human T-cell
virus types I and
lymphotropic virusII;types
SFV,Isimian foamy
and II; SFV, virus;foamy
simian SRV, virus;
simian retroviruses;
SRV, STLV, simian
simian retroviruses; STLV,T-lymphotropic virus; virus;
simian T-lymphotropic SV40,SV40,
simian virusvirus
simian 40. 40.
US Dept Health
US DeptServices,
Human Health Human
Food andServices, Food and Drug Administration.
Drug Administration 23
23
. (Table 1, Gene Therapy 15, 840–848 (2008))
Table reprinted with permission from Springer Nature
Quality control analytical services
• Plasmid QC
◆ Purity & identity: % supercoil (HPLC), sequencing, residual host DNA/RNA/proteins
◆ Safety: sterility, endotoxin, mycoplasma, etc.
• Viral vector QC
◆ Purity and impurities: residual IDX (HPLC), residual host DNA/RNA/proteins
◆ Strength & safety: vector genome titer (qPCR, ddPCR), total particle titer (ELISA),
sterility, mycoplasma, etc.
Please view the promo at vigenebio.com/2021/AugViral vector and plasmid analytical services
MONDAY TUESDAY WEDNESDAY THURSDAY
AUGUST
FRIDAY
2021
SATURDAY SUNDAY
1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30 31
www.vigenebio.comSelected
Table BCMABCMA
1 Selected CAR-T trialstrials.
CAR-T
Institution/ Vector/co-stimulatory domain BCMA positivity requirement No of patients Median prior lines Efficacy Safety
developer (range) ORR*/PFS CRS/ICANS
(months)
NCI γ-retrovirus/CD28 >50% 24 9 (3–13) 81%/7.2 94% (38% grade ≥ 3)/NE
UPenn Lentivirus/4–1BB Not required 25 7 (3–13) Cohort 1 44%/2.2 88% (32% grade ≥ 3)/32%
Cohort 2 20%/1.9 (12% grade ≥ 3)
Cohort 3 64%/4.2
Bb2121 Lentivirus/4-1BB ≥50% in dose escalation, NR in 33 7 (3–14) 85%/11.8 70% (6% grade ≥ 3)/42%
dose expansion 8 (3–23) (3.3% grade ≥ 3)
Bb21217 Lentivirus/4-1BB; PI3K inh during in vivo >50% 22 7 (4–17) 83%/NR 59% (4.5% grade ≥ 3)/22%
expansion (9% grade≥3)
LCAR-B38 M Lentivirus/41-BB Required 57 3 (1–9) 88%/15 90% (grade 3 ≥ 7%)/2%
LCAR-B38M Lentivirus/4-1BB Required 17 4 (3–11) 88%/NR 100% (grade ≥ 3 35%)/NR
Poseida (P-BCMA 101) PiggyBAC/4-1BB Not required 23 6 (3–11) 63%/NR 9.5%/4.8% (grade ≥ 3 4.8%)
JCARH125 Lentivirus/4-1BB Not required 44 7 (3–23) 82%/NR 80% (9% grade ≥ 3)/25%
(grade ≥ 3 7%)
MCARH171 Retrovirus/4-1BB/tEGFR Required 11 6 (4–14) 64%/NR 60% (20% grade ≥ 3)/NR
Han et al Lentivirus/4-1BB/Alpaca VHH Not required 16 10 (NR) 100%/NR NR (12.5% grade ≥ 3)/NR
FCARH143 Lentivirus/4-1BB/tEGFR ≥5% 7 8 (6–11) 100%/NR 86%/0%
CARTITUDE-1 Lentivirus/4-1BB Not required 25 5 (3–16) 91%/NR 80% (8% grade ≥ 3)/12%
(4% grade ≥ 3)
CT053 Lentivirus/4-1BB ≥50% 16 NR 100%/NR 18% (6% grade ≥ 3)/NR
CT103 Lentivirus/4-1BB NR 16 4 (3–5) 100%/NR 100% (37.5% grade ≥ 3)/0%
Cowan et al Lentivirus/4-1BB Required 8 10 (4–23) 100%/NR 100%/70%
GSI (JSMD194)/tEGFR
C-CAR088 Lentivirus/4-1BB NR 3 7 (NR) 100%/NR NR
HRAIN biotechnology γ-retrovirus-4-1BB/tEGFR >5% 17 NR 79%/NR NR
CRS cytokine release syndrome, ICANS immune effector cell associated neurotoxicity syndrome, ORR overall response rate, PFS Progression free survival, BCMA B cell maturation antigen, GSI
CRS cytokine release syndrome, ICANS immune effector cell associated neurotoxicity syndrome, ORR overall response rate, PFS Progression free survival, BCMA B cell
gamma secretase inhibitor, EGFR epidermal growth factor receptor.
maturation antigen, GSI gamma secretase inhibitor, EGFR epidermal growth factor receptor.
*responses
*responsesassessed afterafter
assessed 30 days.
30days.
(Table 1, Bone Marrow Transplantation https://dx.doi.org/10.1038/s41409-020-01023-w (2020))
Table reprinted with permission from Springer Nature
B. Dhakal et al.
Retrovirus production – Research, preclinical, clinical
• Research grade, preclinical, clinical, and commercial GMP retrovirus production
• High titer, purified, ready to use in vitro and animal studies – research grade
• Chromatography based commercial ready retrovirus production – GMP
• High transduction efficiency
Please view the promo at vigenebio.com/2021/SeptRetrovirus production
MONDAY TUESDAY WEDNESDAY
SEPTEMBER
THURSDAY FRIDAY
2021
SATURDAY SUNDAY
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30
www.vigenebio.comConventional ssAAV ScAAV
wtTR ΔTR
2.3 kb
4.6 kb
wtTR
wtTR
4.6 kb 4.6 kb
3ʹ 5ʹ 3ʹ 5ʹ
+
Open Open +
4.6 kb 4.6 kb
3ʹ 5ʹ 3ʹ 5ʹ
–
Open Open –
Graphical representation of the suggested portion of the transgene plasmid to be
used as the template of the probe for dot-blot analysis. The black lines represent
sequences in the bacterial backbone; the blue and red lines represent sequences
of the coding and complementary sequence of the transgene expression cassette,
respectively. (Figure 7, Nature Protocols 1, 1412–1428 (2006))
Figure reprinted with permission from Springer Nature
AAV & LVV plasmid ITR/LTR repair and production service
Plasmid design and cloning (identical ITR vs hybrid)
• Genetic stability study & ITR sequencing analysis with NGS
• AAV plasmid & LVV plasmid clonal screening using different E. coli strains
for stable and reproducible ITR/LTR plasmid production
Please view the promo at vigenebio.com/2021/OctAAV plasmid ITR repair and production service
MONDAY TUESDAY WEDNESDAY THURSDAY
OCTOBER
FRIDAY
2021
SATURDAY SUNDAY
1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
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www.vigenebio.comPackaging and payload Delivery Tropism
Local Global
injection injection All cells Selected cells
Gene X
Virus particle Infected cell Uninfected cell
Genetic access Infectivity and toxicity Transgene expression
Entry Virus
Transgene expression
(susceptibility) Infectivity injection
Healthy
cell
m7G AAA Transgene
expression
(permissivity) Toxicity
Damaged Duration of Decay or
cell expression toxicity
Onset
Key principles for viral-mediated gene transfer in neuroscience. Schematic demonstrating six key principles essential for the
neuroscientist: viral packaging limit (how much nucleic acid a virus particle can carry) and payload (the length and type of genomic
material that can be successfully packaged into a virus particle), delivery methods (local versus global injections), tropism (specificity
of a virus for a given cell type(s)), access (ability of a virus to enter a cell type and express its gene product(s)), infectivity and toxicity
(how efficiently a virus infects a cell and how harmful it is to the cell), and transgene expression dynamics (time course of onset and
persistence of transgene expression). These principles play a key role in determining a neuroscientist’s choice of virus by weighing the
advantages and disadvantages of a given virus. AAA, 3´ poly(A) tail for mRNA; Gene X, a transgene being packaged into a virus particle;
m7G, 7-methylguanosine (5´ cap for mRNA). (Figure 1, Nature Reviews Neuroscience 21, 669–681(2020))
Figure reprinted with permission from Springer Nature
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2D cell factories Microcarrier culture (1) Subculture — Modified surfaces
bead-to-bead transfer
(2) Dissociation
Solid
microcarrier
Degradable
Enzymatic surface
digestion
Engineered substrates
PNIPAM
n
O N
T < 32 °C T > 32 °C
Centrifugal
separation
Bottlenecks Solutions
Process optimization for the expansion of cells and for cell collection from microcarriers. a, Production of clinical lots by using adherent MSCs
in 2D cell-culture plates. Issues with the scaling of costs and labour efficiency make 2D culture unlikely to meet an estimated demand of > 1012
viable cells per year, necessary for treating prevalent adult indications. b, Suspension culture systems for MSCs use microcarriers and stirred tank
bioreactors and are a scalable and sustainable approach for cell expansion at high density. c, Unit operations identified as major bioprocessing
bottlenecks: (1) bead-to-bead transfer for MSC subculturing and expansion; (2) the need for enzymatic digestion and centrifugal separation to
isolate the MSCs from the microcarriers. d, Materials-science innovations in microcarrier substrates can improve product purity, identity and
potency through degradable and temperature (T)-sensitive materials (such as poly(N-isopropylacrylamide), PNIPAM) that remove the need for
additional enzymatic dissociation processes. (Figure 1, Nature Biomedical Engineering 2, 362–376 (2018))
Figure reprinted with permission from Springer Nature
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