APPLICATION NOTE

Send your LNP product development to lightspeedwith the Sunny SuiteIntroduction
Lipid Nanoparticles are changing the face ofmedicine – from rapid responses to infectious
disease, to in-vivo gene editing and targeted protein
replacement therapeutics, LNPs are leading theway. Despite their proven clinical success, LNPs stillpresent challenges to the drug developer, among
them that the material produced in discovery must
be the same as that produced commercially. Asa complex, formulated product, this is not always
easy, as nuances in the formation of the particlescan affect particle size, distribution structure, andin the end, efficacy (Figure 1).
Unchained Labs’ Sunny Suite leads our customers
through the whole process of pharmaceutical productdevelopment – from the first steps of screening large
libraries of formulations, all the way through to themanufacture of clinical materials (Figure 2).

Figure 1: Experimental requirements at the different stages ofdrug development.

Figure 2: Sunny Suite instruments across the drug development stages.

All the method development and changes in process
throughout the lifetime of a product development
project can leave you feeling stuck in the mud on thepath to production, raising costs and eating time.

To beat the process transfer blues, the Sunny Suite
equipment is designed to change with your needs ateach step as the product gets closer to realization,whilst maintaining process consistency in between
each device, making the transition as easy as possible.

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SEND YOUR LNP PRODUCT DEVELOPMENT TO LIGHTSPEED WITH SUNNY SUITE

Surf to seamless scale-up at GMP
with Sunbather
Good Manufacturing Practice (GMP) for clinicalmaterial comes with its own additional burdens– the software has got to be FDA 21 CFR Part 11
compliant for full traceability, the flow path has tobe single-use or clean-in-place (CIP) compatible
to avoid bio-contamination, and all the contact
materials have to be tested for leachables andextractables to make sure there’s no surprise extra
chemicals in there! It’s a high bar to clear, but don’t
worry…

Figure 4: Sunny 100X.

Uniquely, the pumping technology is also retained
from initial screening all the way to clinicalproduction. Therefore, you know exactly what you
are getting from your pump performance, with noadditional process development necessary as you
shift to manufacture (Figure 5).

Sunbather (Figure 3) is Unchained Labs’ solution
to GMP manufacture. Unlike other GMP solutions,Sunbather is designed to require no further
optimization from the method already developedon the Sunshine instrument in continuous
mode, by exactly maintaining the core system
functionalities of each device.

This means there is much less danger of process
hiccups as you move into GMP manufacture, andmakes drastic savings on time, materials andcost (Figure 6).

160

0.5
0.45

140

Made from glass with ultra-smooth channels toavoid material build-up, these are as well-suitedfor running multiple formulations in a screen as
they are for long, continuous production runs.

0.4

S i z e (n m)

1200.35

1000.3

80

0.25
0.2

600.15400.1

2000.05
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31
A li quot Num ber

0Figure 5: Aliquot of a continuous production run on Sunshine
using a Sunny 490 Trident T to optimize particle production
before transfer to GMP.

Figure 3: Sunbather, the GMP-ready LNP producer.

2PDI

This includes our advanced microfluidic mixing
devices, or “Sunnies,” (Figure 4) that let you use thesame mixing conditions across the development
process, including GMP.

SEND YOUR LNP PRODUCT DEVELOPMENT TO LIGHTSPEED WITH SUNNY SUITE

Microfluidic mixing gives excellent
control over your process
throughout development andmanufacture.

Provided with all necessary
traceability documentation forregulatory compliance.

The same super accurate
pumping technology works acrossyour Sunny Suite instruments, sono more tricky transfers.

Excellent service and support,
from installation and training
through to after-care productsupport.

Our highly reliable process means
fewer wasted batches, and saves
time and money.

Combining these benefits means
you get your life-changing
vaccines and therapeutics tomarket faster than ever before.

Figure 6: Sunbather benefits.

Put poor process control in theshade with a Sunny

Sunbather uses a fully replaceable wetted flow-path(with accompanying extractables data provided),comprised of pump valves, syringes and tubing, anda GMP-Sunny of your choice, to eliminate the chancesof batch contamination. Replace the whole flowpath
run-to-run, or clean in place with a cleaning protocol– the choice is yours. The control unit is a touch screen
kiosk perfect for a production environment, with21 CFR Part 11 compliant software for full process
traceability. The software is designed for ease of use,with minimal training necessary (Figure 7).

Our microfluidic Sunnies are more than just simple
mixing channels. Each one is made of chemicallyinert glass, which has a channel surface roughness ofjust ±5 nm, compared to injection molded chips thattypically have values of ±1-3 µm!
Their careful design allows for thorough, controlled
mixing. Various mixing geometries mean your mixing
can be suited to your exact application and the flow
rates that you need for your particular product andyour process.
The Sunny is compatible with each of the SunnySuite systems, and is used with Sunbather alongwith all the tubing, fittings and syringes (for the pumpchannels) needed to run the device whilst minimizing
contamination risk. The flowpaths come with thenecessary documentation for material traceability
and accountability, and Sunbather is available with afull warranty, IQ/OQ and maintenance packages soyou know we’ve got your back.

Figure 7: Sunbather’s easy-to-use software interface.

Just hook up your bioprocessing bags, plug in the flow
rates developed from your Sunshine continuous runs,
and set it going to produce as much nanoparticle
suspension as you need, with no fuss.
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SEND YOUR LNP PRODUCT DEVELOPMENT TO LIGHTSPEED WITH SUNNY SUITE

transitioned the Sunshine to continuous modeand produced 20 mL of mFLuc-LNP suspension.
Finally, the same formulation and conditions wereused with Sunbather, to produce 60 mL of LNPs ina GMP appropriate process.

Reliable results so you can relax
The excellent microfluidic reproducibility means
you know you can go from one development stage
to another with the same excellent control overthe process, so you know your batch will be up toscratch when it’s time to manufacture.

All these samples were analyzed for consistency,
measuring particle size, PDI (Figure 8) and EE%(Figure 9) – showing excellent reproducibility acrossthe sunny suite range, with unbeatable ease andconvenience.

Let’s walk through a process then, using theModerna Spikevax formulation as a commerciallyrelevant example, with FLuc mRNA as the cargo.
Starting with Sunscreen, we’re looking for aformulation that produces stable particles withgood transfection efficiency. In the research
stage, you’ll be running hundreds of different
formulations, which Sunscreen is perfect forwith its high-throughput, fully automated 96well plate format. The reusable microfluidic chipand small sample sizes (from as low as 100 µL/input) help you cut your costs, too! To see what
96 formulations in a day could look like foryou, check out the App Note “Accelerate yourLNP formulation screening with Sunscreen’s
automated microfluidic technology.”

1120

0.9
0.8
0.7

80

0.6
60

0.5

PDI

S i z e (n m )1000.4
400.3
0.2

200.1
00Sunscreen

Sunshine
ProtocolSunshine
Continuous

SunbatherFigure 8: Comparison of LNPs sizes produced on different SunnySuite instruments.

When you are happy with one formulation, (or, as
it may be, 10 or 20 formulations), Sunshine helps
shine a light on the best process to use as you
start to optimize your product. Sunshine allows
rapid formulation and process prototyping, withlarger and more flexible sample volumes, from ~1mL up to fully continuous manufacture. Just selectyour winning screening formulation, run samples
in protocol mode for optimization, then move on tocontinuous mode, to make large volumes of yourmost promising candidates.

10090
80

EE%

7060
50
4030
20
100Sunscreen

Once you’ve done that, it’s just a hop and a skip
to move to GMP manufacture with Sunbather– no optimizing necessary. Just use one of theSunbather Flowpath Packs that matches with yourSunshine configuration and go for it.

Sunshine
ProtocolSunshine
ContinuousSunbatherFigure 9: Comparison of LNP Encapsulation Efficiency (EE%)
produced on different Sunny Suite instruments.

The high-performance continuous syringe pumps willkeep on pushing out material for as long as you needthem to, with consistent, reliable performance. We’ve
produced liters of material here, so you can be surethat Sunbather can get the job done for you.

To show how easy the path to GMP manufacture
can be, we’ve produced the same formulation,at the same conditions, across all of our devices.
Starting with Sunscreen, we collected 800 µL
of mFLuc-LNPs, using the Sunny 490 Trident Tat 12 mL/min. We ran the same formulation onSunshine with a 1.5 mL collection volume, then4SEND YOUR LNP PRODUCT DEVELOPMENT TO LIGHTSPEED WITH SUNNY SUITE

Join the crew

Get ready to breeze through toGMP

As if that wasn’t game-changing enough, UnchainedLabs also helps you free up your quality controlworkflow. Stunner, the first-in-class Rotational Angle
Dynamic Light Scattering (RADLS) system, allows you
to analyze the quality of your material.

From formulation screening to clinical manufacture,
this is GMP made as easy as it can be. Reducing
the time, cost and failure rate of your development
processes, Unchained Labs helps you get the best
version of your product to market as quickly aspossible.

With a 96-well plate format input you can churnthrough samples rapidly, and at only 2 µL per sample,
with a minimum of wasted material. Combinedwith simultaneous total RNA quantification andparticle concentration, there is no better solution toensure your particles are hitting their Critical Quality
Attributes of size, distribution, concentration andRNA content (Figure 8).

Unchained Labs
4747 Willow Rd,Pleasanton, CA 94588
Phone: 1.925.587.9800
Toll-free: 1.800.815.6384
Email: info@unchainedlabs.com© 2024 Unchained Labs. All rights reserved. The UnchainedLabs logo, Sunscreen, Sunshine, Sunbather, Sunny andSunnies are trademarks and/or registered trademarksof Unchained Labs. All other brands or product names
mentioned are trademarks owned by their respective
organizations.

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