Separations,
Journal Year:
2024,
Volume and Issue:
11(12), P. 335 - 335
Published: Nov. 21, 2024
Protein
purification
is
a
crucial
step
for
various
downstream
applications
like
drug
development,
antibody
preparation,
and
structure
determination.
The
constant
pursuit
methods
that
are
more
efficient
cost-effective.
We
propose
novel
approach
using
an
elastin-like
polypeptide
(ELP)
as
aggregation
core
serves
anchor
between
the
beads
in
chromatography
column.
In
this
method,
chilled
sample
containing
[target
protein
type]
fusion
loaded
onto
pre-equilibrated
IMAC
(immobilized
metal
affinity
chromatography)
column
with
low-salt
buffer.
then
washed
warm
buffer
high
salt
to
remove
impurities.
Here,
key
involves
warming
above
ELP’s
transition
temperature
(Tt),
which
triggers
its
aggregation.
This
expected
trap
target
tightly
beads.
Subsequently,
harsh
wash
imidazole
can
be
applied
even
persistent
contaminants,
achieving
purity.
Finally,
lowered,
cold,
introduced
reverse
elute
purified
protein.
method
has
potential
eliminate
need
sophisticated
systems
while
still
PLoS ONE,
Journal Year:
2025,
Volume and Issue:
20(4), P. e0321094 - e0321094
Published: April 29, 2025
The
prolonged
persistence
of
extracellular
chromatin
and
DNA
is
a
salient
feature
diseases
like
cystic
fibrosis,
systemic
lupus
erythematosus
COVID-19
associated
microangiopathy.
Since
deoxyribonuclease
I
(DNase1)
major
endonuclease
involved
in
DNA-related
waste
disposal,
recombinant
DNase1
an
important
therapeutic
biologic.
Recently
we
described
the
production
murine
(rmDNase1)
Pichia
pastoris
by
employing
α-mating
factor
prepro
signal
peptide
(αMF-SP)
method,
which
now
applied
to
express
human
DNASE1
(rhDNASE1).
In
addition
impaired
cleavage
αMF
pro-peptide,
also
detected
previously
for
mDNase1,
expression
hDNASE1
resulted
70–80
times
lower
yield
although
both
orthologues
share
high
structural
functional
homology.
Using
mDNase1
as
guideline,
were
able
increase
fourfold
optimizing
parameters
nutrients,
cultivation
temperature,
methanol
supply,
codon
usage.
addition,
post-translational
import
into
rough
endoplasmic
reticulum
(rER)
was
changed
co-translational
(SP)
α-subunit
Oligosaccharyltransferase
complex
(Ost1)
from
Saccharomyces
cerevisiae
.
These
improvements
purification
~
8
mg
pure
mature
rmDNase1
0.4
rhDNASE1
per
Liter
medium
culture
with
cell
density
OD
600
=
40
24
hours.
As
main
cause
difference,
assume
varying
folding
abilities
reach
native
conformation,
induce
elevated
unproductive
unfolded
protein
response
within
rER
during
expression.
Concerning
functionality,
expressed
P.
comparable
Pulmozyme®,
i.e.
produced
Chinese
hamster
ovary
(CHO)
cells
Roche
-
Genentech.
With
respect
biochemical
effectivity,
superior
due
its
higher
specific
activity
presence
Ca
2
+
/Mg
inhibition
monomeric
actin.
Biotechnology for Biofuels and Bioproducts,
Journal Year:
2025,
Volume and Issue:
18(1)
Published: March 15, 2025
The
growing
demand
for
sustainable
and
eco-friendly
alternatives
in
bioprocessing,
healthcare,
manufacturing
has
stimulated
significant
interest
Bacillus
subtilis
surface
display
technology.
This
innovative
platform,
leveraging
both
spore
vegetative
cell
forms,
provides
exceptional
versatility
a
wide
spectrum
of
applications,
spanning
from
green
technologies
to
advanced
biomedical
innovations.
robustness
spores
the
metabolic
activity
cells
enable
efficient
enzyme
immobilization,
biocatalysis,
biosensor
development,
facilitating
bioremediation,
pollutant
degradation,
renewable
energy
generation.
Additionally,
B.
systems
have
demonstrated
remarkable
potential
vaccine
development
drug
delivery,
offering
cost-effective,
scalable,
environmentally
alternative
traditional
methods.
These
can
effectively
present
antigens
or
therapeutic
molecules,
enabling
targeted
delivery
robust
immune
responses.
review
explores
recent
advancements,
challenges,
opportunities
harnessing
technology
biomanufacturing,
innovations,
transformative
emphasizing
its
role
addressing
pressing
global
challenges
environmental
sustainability
healthcare.
