Genetic Engineering News article on transfection of DNA Vaccines. Several DNA Vaccines experts, including Althea's Magda Marquet, share their perspectives on DNA vaccine delivery and the future of theis great technology.

1. Volume 33, number 1 Tools | Technologies | Trends January 1, 2013
CHRONIC LYMPHOCYTIC LEUKEMIA
HEPATOCELLULAR CARCINOMA
ALZHEIMERS OR ALZHEIMER'S
EPITHELIAL*MESENCHYMAL
ACUTE MEYLOID LEUKEMIA
COMPANION DIAGNOSTIC
DEAF OR HEARING LOSS
HODGKINS LYMPHOMA
COLORECTAL CANCER
Biobusiness
BURKITT LYMPHOMA
ADENOCARCINOMA
HCV OR HEPATITIS C
B-CELL LYMPHOMA
BLADDER CANCER
GASTRIC CANCER
HEPATIC CANCER
HEMATOPOIETIC
INFLAMMATION
HEMATOPOIESIS
BONE MARROW
COLON CANCER
GLIOBLASTOMA
HEART DISEASE
ANGIOGENESIS
LUNG CANCER
Breast Cancer
MELANOMA
METASTASIS
BIOMARKER
LEUKEMIA
EXOSOME
DIABETES
EMT
MARKER
14-3-3
ALPHA-FETOPROTEIN
BLADDER TUMOR ANITGEN
0
0
1 25
170
0
30
0
0
4
17
0 0
1
1
34
1
4
37
13 8
4
4
68
0
39
47
0
0
0
0
2
0
0
8
9
0
8
12
0
Cancer 0
0
0
3
0
1
22
38
0
1
2
0
2
0
5 4
0
0
110
6
0 0
1
1
6
2 15
0
0
2
0
8
20
0
4
2
0
0
10
0
10
2849
0
0
0
0
11
17
0
19
13
0
28
31
0
6
9
0
14
18 4
0
Biomarker
B-TYPE NATRIURETIC PEPTIDE 0 0 0 4 0 0 235 4 6 0 0 0 0 0 0 46 0 0 0 0 0 4 0 1 0 0 0 0 31 0 9 0 0
CA 125 0 14 3 0 9 2 7 85 7 56 1 2 2 12 0 0 1 0 0 1 9 0 2 0 5 1 0 6 1 12 5 41 3 53
CA 15_3 0 20 0 0 0 2 21 1 435 0 0 1 6 0 0 0 0 0 0 2 0 0 0 2 0 0 1 0 0 1 7 1 33
Hotspot
CA 19-9 1 18 1 0 3 0 29 39 4 15 0 0 15 14 7 0 0 9 0 0 0 10 3 0 5 0 4 0 0 22 0 5 3 27 2 64
CD98 0 1 0 1 0 0 0 0 4 0 0 0 0 0 0 1 0 0 0 0 1 0 2 2 0 0 0 0 5 2 5 0 1
CEA 0 805 1 15 1 58 152 48 627 0 1 3 13 12 6 6 0 3 27 0 1 4 4 59 5 7 0 17 2 6 88 0 31 14 58 8 51 449
Analysis
FASCIN 0 22 1 4 2 3 6 2 11 0 0 2 6 0 2 0 2 3 1 3 5 0 0 3 0 0 4 0 3 1 5 5 20
HER2_NEU 1 264 1 78 1 53 241 47 54 4 5 0 2 21 46 4 1 7 14 13 7 153 30 3 0 14 0 0 19 0 20 33 231 44 287
IGF-11 0 32 9 10 0 2 14 14 13 7 0 0 30 27 0 0 92 0 0 0 10 6 9 3 11 0 1 80 0 14 21 23 6 14
LEPTIN 3 24 30 71 2 1 119 62 19 5 0 1 32 39 0 1 12 79 1 2 1 12 5 47 15 22 1 0 25 0 52 7 43 18 16 23
OSTEOPONTIN 2 26 4 55 2 1 10 1 2 12 10 7 0 0 9 23 0 0 49 0 6 0 18 9 14 4 31 1 1 65 0 154 19 43 43 19 4
PROLACTIN 2 55 34 54 1 1 56 24 668 1 0 2 9 0 36 19 5 3 2 1 1 3 11 6 33 6 0 7 0 38 36 7 3 28
11
PSA 1 56 9 9 51 1 30 299 39 99 0 1 6 14 0 4 40 9 6 2 3 2 6 1 4 2 0 6 0 10 2 23 26 6 2 13
THYROGLOBULIN 0 35 4 3 1 1 10 12 9 1 0 2 2 0 14 116 0 2 0 0 0 35 0 1 1 0 5 0 10 12 5 7 10 0
TROPONIN I 1 0 0 15 1 0 2 10 44 14 0 0 0 1 0 0 20 1 0 0 0 0 2 0 7 0 0 0 0 60 4 1 1 0
Drug Discovery
Novel Aldevron’s Genetic
Immunization and
Strategies for Antibody (GIA™)
technology is used
Cell-Based to test new DNA
Assays vaccine techniques
in animal models.
14
OMICS
Genomic
Tackling Transfection Tasks
Neil McKenna, Ph.D.
Analysis Drives
Biomarker The discovery in the early 1990s that administration in mice of plasmid DNA encoding
Discovery both viral and nonviral antigens induced antibody responses held out the prospect of
achieving broad immunogenicity using DNA vaccines without the safety issues associated
26 with a replicating pathogen.
Despite this promise, the efficacy of first-generation DNA vaccines against HIV, HPV, and hepatitis was compromised by low an-
Bioprocessing tibody titers and sporadic immune cell responses. In addition, DNA vaccines face broader challenges encountered by other classes of
vaccines such as manufacturing, scale-up, and purification, as well as vaccine resistance.
A number of companies are developing processes and technical platforms designed see page 22
Twisting and
Turning for
Lab Automation Sparks Creativity
Better Protein
Expression
30
Kate Marusina, Ph.D.
Translational Just two decades ago we were only beginning to recog-
Medicine Necessary nize the potential of automated technologies to enhance
Liaisons: throughput in drug discovery research. Today, it is difficult
CROs Gain to imagine a modern laboratory without robotic equipment.
Clout & “We see continuous trends toward increased reliability of robots, partially driven by the
Responsibility Primary mouse embryonic fibroblast strain introduction of new types of motors,” reports Malcolm Crook, Ph.D., CTO, Process Analysis
DBA/2J cells imaged using Olympus 40X/0.9NA & Automation (PAA). “Another trend is toward smaller targeted systems, that still flexibly
COMPATIBILITY
36 SENSING
air objective lens on the BD Pathway 435 accommodate peripherals as needed.”
High Content Imager. Cells were labeled PAA will be presenting the “automate.it harmony,” its user interface for software inte-
with Hoechst 33342 (blue), YoYo-1 (green),
gration, at the Society for Laboratory Automation
ONE TOUCH
TEACHING
www.genengnews.com
MitoTracker Orange (red), and Cytochrome C –
SCREENING
AlexaFluor 647 (cyan).
RELIABILITY
Hamner Institute and Screening (SLAS) meeting later this month. see page 16
CAPABILITY MULTISPAN HIGH THROUGHPUT OPTIMIZATION
CAPABILITY
HOW EXTRAORDINARY HAPPENS IntroducIng the Agilent encore
MultispAn liquid HAndling systeM
www.agilent.com/lifesciences/see-it-happen
CONFIDENCE
MODULARITY FLEXIBILITY SPEED CONTROL
ACCURACY

2. OMICS
Transfection Continued from page 1
to circumvent these issues, with particular across the cell membrane to avoid degrada-
emphasis on improving the efficiency of de- tion by endonucleases.
livery and uptake of vaccines by target cells, Inovio’s approach in this area includes
referred to as transfection, which can be con- informatically assisted optimization of the
founded by differences in tissue and cell type coding sequence to expand immunogenici-
accessibility between individuals, in addition ty across multiple targets and boost expres-
to other factors. sion levels of immunogens in target cells,
Many of these companies were present and to improve the delivery of the plasmid
at last month’s DNA Vaccines conference in DNA into host cells to minimize exposure
San Diego. The meeting was sponsored by to extracellular nucleases. The development
International Society of DNA Vaccines and of one of Inovio’s candidate high-grade
organized by BioConferences International, cervical dysplasia vaccines, VGX-3100,
a Mary Ann Liebert company. encoding the HPV16 and HPV18 E6/E7
“Vaccines have saved more lives than antigens, incorporated a combination of
any other invention in human history”, ex- these features to increase vaccine antigen
plained J. Joseph Kim, Ph.D., president and immune potency.
CEO of Inovio Pharmaceuticals. “Conven- “The VGX-3100 vaccine was developed
tional vaccines have been successfully used using our SynCon platform and includes
against the lower hanging fruit in terms of highly efficient leader and Kozak sequenc-
disease, and more complex diseases such es,” said Dr. Kim. “We also introduced an
as cancer represent both opportunities and endoproteolytic cleavage site to improve
challenges for DNA vaccines. protein folding and cytotoxic T lymphocyte
According to Dr. Kim, one of the biggest processing.” Vaccine delivery using electroporation (>10–100x enhancement in immune responses). Inovio
hurdles for DNA vaccines has been induc- In addition, Inovio has developed an elec-
ing sufficiently high immune responses for troporation-based delivery system that, ac-
effective vaccination. “There are two issues cording to Dr. Kim, allows for more efficient siently realign into a more porous state,” he Looking to the future, “Electroporation is
at stake,” he said. “The first is to make plas- and safe targeting of the antigenic sequence explained, adding that electroporation is the not a static technology,” continued Dr. Kim.
mids more ‘people-friendly’ to optimize their to the target cell. “A low voltage electrical most efficient and safe mode of delivery of “We are developing new devices, referred
expression once in the cell, and the second field is applied at the site of vaccine injec- nucleic acids, requiring no additional chemi- to as surface electroporators, that sit on the
is to optimize the rapid transfer of the DNA tion, causing the cell membranes to tran- cals, preservatives, or adjuvants. See Transfection on page 24
NEWS Genomics & Proteomics
> CLC bio Contributes processes—how, when, and why are cer- expression using flow cytometry and mul- complete BioCyc Pathway Database Col-
Expertise to STATegra tain genes switched off or expressed?” tiplex assays, solidifying the connection lection. The integrated package will allow
CLC bio is the latest company to par- states Michael Lappe, Ph.D., senior bioin- between the genotype and phenotype of scientists to access, combine, visualize, and
ticipate in the European FP7 project, STAT- formatics scientist at CLC bio. “We aim to interest. These types of connections pro- analyze biological datasets across multiple
egra. It will receive $1.3 million out of the leverage a framework which makes this vide confidence to preclinical and clinical “omics” experiments (genomics, transcrip-
total $7.8 million budget for its efforts. The extremely complex network biology un- decision-makers regarding drug safety and tomics, proteomics, and metabolomics).
STATegra project aims to develop an inte- derstandable. Ultimately our tools will fa- efficacy, says Christina Shasserre, vp of dis- The new software package will enable re-
gral analysis platform for different omics cilitate the translation of massive amounts covery and development. searchers to display biological pathway data
data that is capable of providing a more ef- of data into useful insights that can be ap- generated by Agilent’s suite of instruments,
ficient use of genomics technologies. This plied in clinical settings.” > Agilent and SRI Combine software, and reagents. Agilent products
project will generate and integrate data Within the overall project, methods Technologies for New Offering include microarrays and next-generation se-
obtained in proteomic, metabolomics, and for the validation and design of follow-up Agilent Technologies and SRI Interna- quencing technology, as well as LC/MS, GC/
epigenomic experiments. studies are implemented. Utilizing and val- tional signed a licensing agreement to of- MS, ICP/MS, and NMR systems.
“One of the big challenges today is go- idating existing datasets, STATegra is also fer laboratory research customers a pack-
ing past the raw sequence data and getting going to feed its experimental results back age that combines Agilent’s latest release > Cancer Genome Consortium
to grips with the complexity of epigenetic into the public domain. of GeneSpring GX, GeneSpring NGS, Mass Expands China Research
Profiler Professional, and Pathway Archi- The International Cancer Genome Con-
> EMD Millipore Institutes tect version 12.5 with SRI International’s sortium is expanding its research in China,
Personalized Medicine Services where new projects seeking genomic factors
EMD Millipore’s discovery and devel- in four cancer types common to the world’s
opment solutions business is now offer- most populous nation will be launched.
ing genomic biomarker services. Com- The projects—focusing on colorectal,
pany officials say the purpose is to aid esophageal, liver, and nasopharyngeal
drug companies in targeting patient sub- cancers—join the consortium’s two-year-
groups for improved clinical outcomes, old and still-ongoing Chinese project fo-
understanding drug safety and efficacy, cused on gastric cancer, which made new
and characterizing tumors. Full-scale version of methionine metabolism data from sequenced tumors public.
Researchers at EMD Millipore recently pathway for Saccharomyces C., using BioCyc The new projects are expected to take
content and rendered visually in “simple” mode
profiled transcription in gefitinib-treated less time than the typical four years, since
Rendering of the nucleosome (DNA blue, in MPP/Pathway Architect 12.5; also shows
histone proteins green) highlighting positions non-small cell lung cancer cells by qRT-PCR. average, normalized differential abundances the consortium began collecting tissue
of known epigenetic marks in spherical The genotypic and mRNA expression data for four conditions in the HeatStrips for some samples from patients with the diseases
representation. CLC Bio were then cross-validated with protein of the metabolites. over the past two years. n
22 | January 1, 2013 | genengnews.com | Genetic Engineering & Biotechnology News