FIELD OF TI-IE INVENTION
The present invention relates to biomanufacturing apparatus, for example for cell culturing.
s in particular, the invention relates to bioreactor apparatus in the form of single instruments, and
plural instruments arranged into a biomanufacturing system for optimising the usage of laboratory
and cell culturing space for biomanufacturing.
BACKGROUND OF THE JNVENTJON
10 Cell culture, for example the culture of mammalian, bacterial or fungal cells, may be
carried out to harvest the living cells for therapeutic purposes and/or to harvest biomolecuies, such
as proteins or chemicals (e.g. pharmaceuticals) produced by the cells. As used herein, the term
"biomolecule" can mean any molecule, such as a protein, peptide, nucleic acid, metabolite,
antigen, chemical or biopharmaceutical that is produced by a cell or a virus. Herein, the term
1s biomanufacturing is intended to encompass the culturing or multiplication of cells, and the
production of biomolecules. The term bioreactor is intended to encompass a generally enclosed
volume capable of being used for biomanufaeturiug.
The cells are generally grown in large scale (10,000 to 25,000 litre capacity) bioreactors
which are sterilisable vessels designed to provide the necessary nutrients and environmental
20 conditions required for cell growth and expansion. Conventional bioreactors have glass or metal
growth chambers which can be sterilized and then inoculated with selected cells for subsequent
culture and expansion. Media within the'growth chambers are often agitated or stirred by the use
of mechanical or magnetic impellers to improve aeration, nutrient dispersal and waste removal.
In recent years, there has been a move towards 'single use' bioreaetors which offer smaller
25 batch sizes, greater production flexibility, ease of use, reduced capital cost investment and reduced
risk of cross-contamination. These systems can also improve the efficiency of aeration, feeding
and waste removal to increase cell densities and product yields. Examples include WAVE TM bags
(GE Healthcare) mounted on rocking platforms for mixing, to the introduction of stirred-tank
single-use vessels such as those available from Xcellerex Inc (GE Healthcare). With the advent of
30 'personalised medicine', autologous cell therapies requiring many small batches of cells to treat
patients with unique cell therapies has become important.
Manufacturing facilities, such as tissue culture laboratories, for the production of cells and
biomolecules, have traditionally been custom designed and carried out in clean environmcnts to
reduce the risk of contamination. Such facilities are costly to run and maintain and also to modify
if priorities or work demands change. Work stations for maintaining or harvesting the cells within
5 the bioreactors require a specific 'footprint' which occupies a significant floor space in the culture
laboratory. As the workstations spend much of their time unattended, while the cells are growing
in the bioreactors, the laboratory space is not efficiently or effectively used.
An improvement is proposed in WO 2014122307, wherein the laboratory space required
for cell culture is reduced by the provision of customised workstations and storage bays for
10 bioreactors, on which, conventional WAVE type bioreactors and ancillary equipment can be
supported. Large supporting frameworks are required for that equipment.
US6475776 is an example of an incubator for cell culture dishes, which has a single
incubator housing and multiple shelves, however this type of equipment is not suitable for housing
bioreactors.
15 What is needed is the ability to stack multiple bioreactors one on top of another, closely
spaced side by side, in a system that is simple to load, operate and maintain. Ideally such
bioreactors should be capable of tradition fed batch manufacturing where cells are cultured
typically over 7 to 21 days, as well as perfusion type manufacturing where cells can be cultured for
longer periods, but waste products are continually or regularly removed, and biomolecules may be
20 harvested. On top of that, one of the important parameters that needs to be is measured during the
entire cell expansion process is the weight of the cell culture. This parameter serves as an input for
various application steps like step-wise' cell expansion, continuous cell expausion, media fill,
perfusion flow, calibration of feed & harvest pumps, temperature control and pH control. It is
therefore a very important requirement for the weight measurement system to have a very good
25 accuracy and sensitivity specification. A cell culture instrument which has a weight measuring
load cells mounted on each foot of the instrument is known commercially. This is not an ideal way
of mounting the load cells because the entire weight of the instrument rests on the load cells.
Therefore, the dead weight acting on the load cells are very high (depending on the instrument
weigh) which affects the overall accuracy and sensitivity. Also since the load cells are mounted on
30 the instrument feet, there is a significant change in the reading of the load cells when they are
subjected to even a slight side force. This is not acceptable sincc it results in error in the load cell
readings.
An ideal weight measurement system is therefore one which carries as little dead weight as
possible and is also isolated from the instrument feet so that there is no impact on the readings
5 when there is a disturbance to the instrument, and instruments can then be stacked one on top of
another.
SUMMARY OF THE INVENTION
The invention provides an arrangement according to claim 1 having preferred features defined by
claims dependent on claim 1.
10
The invention extends to any combination of features disclosed herein, whether or not such a
combination is mentioned explicitly herein. Further, where two or more features are mentioned in
combination, it is intended that such features may be claimed separately without extending the
scope of the invention.
15
BRIEF DESCRIPTION OF THE DRAWINGS
The invention can be put into effect in numerous ways, illustrative embodiments of which are
described below with reference to the drawings, wherein:
Figure la shows a pictorial view of an embodiment of biomanufacturing apparatus;
20 Figure lb shows the apparatus of Fig la stacked to form a biomanufacturing system 2;
Figure 2 shows a different pictorial view of the apparatus shown in Fig 1;
Figure 3 shows another pictorial view df the apparatus shown in Fig 1, including a bioreactor
loaded inside the apparatus;
Figures 4 and 5 show two pictorial views of a further embodiment of biomanufacturing apparatus,
25 in different configurations;
Figures 6a, 6b, 6c and 6d show a partial sectional view of the apparatus shown in Figs land 2;
Figure 7 shows an enlarged partial view of the apparatus shown in Figures 1 and 2;
Figure 7a shows an exploded view of the apparatus shown in Figure 7;
Figure 8 shows a sectional plan view of the apparatus shown in Figs 1 and 2; and
30 Figure 9 shows a schematic representation of ihe functioning of the apparatus shown in Figs 1 and
2.
The invention, together with its objects and the advantages thereof, may he understood better by
reference to the following description talten in conjunction with the accompanying drawings, in
which like reference numerals identify like elements in the Figures.
5
Referring to Figure la there is shown biomanufacturing apparatus 1 including a generally selfcontained
instrument 10 which includes a generally cuboid or box-shaped housing 20 having
generally flat upper and bottom sides 22 and 24. The bottom side includes four adjustable height
feet 26, only two of which are visible in Figure la. The box shaped housing allows stacking of
10 plural instruments to form a biomanufacturing system. In practice, for convenience, the stack will
be two or three high on a henchtop 5, as schematically illustrated in Figure lb, although there is no
reason why the stack could not be higher. The instrument also includes a door 25, shown open and
cut away for in order to shown the remaining parts of the instrument more clearly. The door is
hinged at hinges 28 to the front vertical edge of the housing, so that it opens about a vertical hinge
15 axis to expose or enclose an insulated chamber 30 inside the housing 20. The chamber 30 is sealed
when the door is closed by an elastomeric seal 32 extending around the whole periphery of the
inner face of the door and cooperating with a seal face 3 1 extending in a complementary manner
around the front edges of the housing 20. No light enters the chamber 30 when the door 25 is
closed. This negates light effects on the cell culture.
20
The chamber 30 has a main chamber 35 and an antechamber 33 leading to the main chamber 35.
The main chamber includes a bioreactor tray 40, supported by a rocking tray support 45 described
in more detail below. The rocking mechanism is protected by a cover plate 21. The antechamber
33 includes a panel 34 supporting two peristaltic pumps only the fluid handling heads 48 and 49 of
25 which extend into the antechamber 33, the electrical parts of which are behind the panel 34. The
panel also includes connections 43 described in more detail below. The antechamber 33 includes
openings 46 defining a route for conduits extending to an external storage area which includes a
bag hanging rack 50.
30 Figure 2 is a different view of the instrument 10 shown in Figure 1, with the door 25 and bag rack
removed 50, in order to show the remaining parts of the instrument more clearly.
Figure 3 shows the instrument 10 of figures 1 and 2, but loaded with a bioreactor 100, in this
instance, in the form of a flexible bag 100, as well as various paths linking the bioreactor to the
instrument, including: a fluid supply conduit 102 feeding the bioreactor with a known mixture of
5 fluids to promote cell growth via the peristaltic pump head 48, a fluid removal conduit 104 for
drawing off fluids from the reactor for the purpose of removing waste components expressed by
cells in the bioreactor via a filter incorporated in the bag 100 and via the peristaltic pump head 49;
a gas feed conduit 106; and paths, for example electrically conductive paths 106, 108 and 110 for
example electrical wires, for various sensors within or adjacent the bioreactor, for example a pH
lo sensor, and a dissolved oxygen (DO) sensor. The conduits and paths can be kept in place by one or
more hangers 23.
Figures 4 and 5 show an embodiment of the instrument 10 including the door 25. The tray 40 in
this embodiment is removable from the tray support 45 by sliding motion and can rested on a
I5 collapsible stand 120, in turn hung on the hinged door 25. In use, the door 25 can be opened, the
I stand 120 can be dropped down, and the tray 40 (without or without a bioreactor in place) can be
I
I slid away from the support 45 and manually moved onto the stand. It will he noted that the tray 40
has an open mid-section. This open section accommodates a bioreactor, which has clips that clip
I
onto the tray 40 sides so that the bioreactor does not fall through the middle of the tray. Returning
20 the tray full or empty back into the chamber 30, allows the frame 120 to be folded away and the
door 25 to be closed shut.
I
Figures 6a, 6b, 6c and 6d each show a sectional view of the main chamber 35 illustrated in Figures
1 to 3, and the components housed therein. Those components include the removable tray 40 and
25 the rocking tray support 45. The tray support 45 is formed from an electrically heated plate 42
which is in direct contact with the bottom of a bioreactor in use, a pivotable plate holder 44 which
releasably holds the heated plate and an electrical stepper motor driving rocking mechanism 47
which moves the plate holder 44 back and forth about a pivot axis P below the tray 40 through a
predefined angle of about 25-35 degrees. The support 45 is controllable in use so that it stops in
30 any position, but in particular in the forward slopping position shown in Fig 6b, which enables the
tray 40 and plate 42 to be slid forward together whilst the plate holder 44 stays in position, to a
new position as illustrated in Figure 6c, where the tray is more readily accessible for loading or
unloading rather than having to remove it as shown in the embodiment of Figure 4 and 5. In the
positon shown in Figure 6c the conduits and paths between the bioreactor and the instrument, as
mentioned above, can be connected or disconnected more easily. The tray 40 and plate 42 can be
5 removed completely as shown in Fig 6d, f o ~ex ample, for cleaning purposes. A cover plate 21
protects the motor and other electrical parts.
Figure 7 shows the rocking mechanism in more detail view from the front, door, side of the
instrument looking into the main chamber 35 with the cover plate 21 removed. A stepper motor 5 1
10 of the rocking mechanism 47 is shown as well as a reduction pinion gear pair 52 driven by the
stepper motor and driving the plate support 44 to rotate back and forth. In this view a load sensor,
in the form of a load cell 41 is visible which in use is used to measure the quantity of fluid added
or removed from the bioreactor. and cell culture control.
15 Figure 7a shows the features of Figure 7 but exploded. The mass change measurement system
consists of a single load cell 41 mounted to support the cell bag tray 40 and the mechanism 47 that
rocks that tray 40. The load cell 41 is a mechanical strain sensor which changes resistance with
strain. However other strain sensor could be used, for example optical strain sensors. Since only
the drive assembly and the rocking platform are mounted over the load cell, the dead load is
20 significantly reduced. The load cell is completed isolated from the exterior and hence is not
affected by any side-forces that act on the apparatus. In use the tray 40 can come to rest in a
horizontal position as shown in Fig 7, Hefore a load measurement is taken. However, it is also
envisaged that 'in use' measurements can be taken, i.e. measurements taken whilst the tray 40 is
rocking. In that latter case an average weight can be determined, and compared with previous
25 averages, to obtain a measurement of weight increase or decrease. The term 'average' is intended
to encompass arithmetic mean, median, mode, range or aggregate loading.
With this weight measurement arrangement, similar apparatus can be stacked one on top of
another. This stacking would not be possible if the load cells are mounted on the apparatus feet
30 because then the apparatus at the bottom of the stack will be measuring the weight of the upper
apparatus also. Another advantage of having the load cell isolated from the apparatus feet is that
the instrument need not be levelled each time before use. In the current bioreactors, the uscr
spends considerable time and effort in levelling the instrument. The wcight measurement system
of the present embodiment rests on the perfectly machined surface and always sees 100% of the
load mounted on it. In traditional bioreactors, each load cell sees a different percentage of the
5 entire instrument weight and the user has to level the instrument so that the percentage of load
seen by each load cell is between 25%-30%. A major advantage is however the improved accuracy
and sensitivity of the present single load cell configuration. For each additional load cell the
accuracy gets poorer by a factor of -\in where n is the number of load cells. The accuracy of the
single load cell configuration is therefore theoretically better than a four load cell configuration by
lo a factor of 2. This is a key benefit when the bioreactor is used for a low volume cell expansion
process.
The modular tray design and a tray eject feature of the apparatus is more fully described in copending
application lN201611015089 filed 29th April 2016. During the entire cell expansion
15 process, there is a need to take daily samples of the cell culture to monitor the progress of the cell
expansion. For taking samples, the instrument door is opened to access the cell bag on the tray.
The tray comes to a stop in an inclined position which ensures that the contents inside the cell bag
come near the sampling port of the cell bag by gravity. When the sampling is carried out by the
user, there is a chance that some weight is transferred on to the rocking platform and therefore to
20 the load cell. To prevent any damage to the load cell due to overloading, the weight measurement
system can have a load cell overload protection which supports the ejected tray or an inclined tray
at the bottom and the load cell is isolated from taking any load at these conditions of the tray. In
other words, when the tray 40 is fully inclined as shown in Fig6b, that arrangement can be
modified such that the lowermost edge of tray 40 can rest on the floor of the chamber 35. The
25 apparatus may also have an alarm for overload protection for any unforeseen circumstances at any
point of time during the entire cell expansion process. The weight measurement system also has an
auto alignment feature to receive the ejected tray back into the correct position to prevent any
misalignment. The user only needs to push thc ejected tray back till an audible click is heard
conveying that the tray is aligned and in position. The user can thus be assured that the tray is safe
30 to start rocking.
Figure 8 shows a sectional view through the instrument 10 looking down such that the main
chamber 35 is visible having a depth D from front to baclc, as well as the antechamber 33, which
has a much shallower depth d. In the remaining region 36 of the housing is separated from the
chambers 35/33 and encloses electrical and electronic control components which are lcept way
5 from possible leaks from the bioreactor and can be kept at lower temperature than the main
chamber, so that electrical parts will have a longer life. In addition, clcaning of the electrical parts
can be avoided because they are separated from the chambers 35/33. In more detail, those
electrical/electronic components include a power supply 37, a perfusion gas supply control unit 38,
a control circuit board 39, a chamber air heater 53, pump head 48/49 drive motors 54/58, a single
10 board computer 55 and various connecting wires and conduits not shown.
Figure 9 shows schematic block diagram of the functioning of the instrument 10, with references
relating to the physical components mentioned above and illustrated in the previous Figures. In use
the flexible bag bioreactor 100 (cell bag) is preferred, and is loaded into the chamber 30 as detailed
15 above. Connections 43 are made and the door 25 is closed. The tray 42, in this embodiment
includes a bar code reader 56, to reader a bar code from ihe bag and relay the identity of the bag to
a controller 39/55. Other identification means are possible, for example an RFID transducer could
be used, embedded in the cell bag 100. The identity of the bag will determine the appropriate cell
culture regime, and additional, external information can be sought by the controller via a system
20 controller 60, for example the target cell density required. Having determined the appropriate cell
culture regime, the controller will, typically, control the temperature external to the bag, and
optimise the parameters inside the bag. 'fhese parameters will vary during the cell culture period,
i.e. over a period of up to 28 days, but typically 7 to 21 days. Thus the controller will monitor and
adjust the internal pH of the cell culture, the dissolved oxygen content of the fluid in the bag, the
25 weight of the bag to dcterrnine the amount of fresh fluid introduced and the amount of waste fluid
withdrawn from the bag. Sampling of these parameters and the cell density is performed
automatically. A continuous perfusion regime is preferred although other known regimes, such as
a fed batch regime could be used. Conveniently, a display 57 is incorporated into the door 25, and
the door includes a window which is darkened to reduce light entering the chamber or has a
30 shutter, openable to view the chamber 30 through the window, but closable to reduce or exclude
light in normal operation of the instrument.
In use the instrument will function as a stand-alone system using the display 57 to output status
information, along with other stand-alone instruments where plural instruments are employed,
meaning that no external control is required for the operation of the instrument or instruments.
5 However, it is possible that the system controller 60 can be used, will function either to simply
supply information relating to the requirements of the ccll bag loaded in the instrument, or
additionally monitor plural instruments, or with suitable software, to monitor and control each
instrument, so that internal instrument control is dominant. The then subordinate controller 39155
of each instrument can take back instrument control if communication with the system controller is
10 lost. The communication between the instruments and the system controller is preferably a system
BUS link for example a universal serial bus of know configuration, but a wireless link is possible,
for example as specified by IEEE802.11 protocols operating at 0.9 to 60 GHz. It is envisaged that
each instrument will be automatically recognised by software running on the system controller,
without the need for any user input.
15
Once the cell culture is complete, as determined by sampling and or cell bag weight, it is removed
from the instrument and used for its intended purpose, for example autologous cell therapy.
Where it is the biomolecules produced by cultured cells that is of interest these can be removed
when the cell bag is emptied, or they can he removed from the filtrate extracted from the bag
20 during culturing. The chamber 30 is easily cleaned ready for the next bag to be introduced, with
minimal down-time. Thus it is apparent that the instrument described above allows convenient
loading and unloading of disposable biorkactors, and can be closely spaced in staclted rows so that
the density of instrumcnts is about 4 to 6 per metre squared when viewed from the instruments'
front faces. A typical bioreactor 100 for use with the instrument 10, will be small by present day
25 standards, i.e. approximately 50 millilitres and 2500 millilitres, and so the system described above
is a small scale system, having multiple cell culture instruments, which are each readily accessible
and controllable, and optimise the available space.
Although embodiments have been described and illustrated, it will be apparent to the skilled
30 addressee that additions, omissions and modifications are possible to those embodiments without
departing from the scope of the invention claimed.

CLAIMS
1. Biornanufacturing apparatus (1) comprising a housing (20) including top (22) and bottom
(24) faces which allow stacking of plural housings, an access door (25) at a front side of the
housing, a substantially enclosed bioreactor chamber (30) inside the housing accessible via the
5 door, and a further substantially enclosed region (36) inside the housing containing electrical parts
and/or electronic control components, the chamber (30) including: a tray (40) for supporting a
bioreactor, a tray support (45) including a mechanism (44,47) for rocking the tray in use, the tray
support further including a load cell (41) to determine changes in the mass load on the tray..
10 2. Biornanufacturing apparatus as claimed in claim 1, wherein the tray support further
includes a sliding portion (42) for sliding the tray at least partially out ofthe chamber via the open
door, the sliding portion being arranged such that when inclined by the mechanism for rocking, a
portion of the mass load on the tray is reacted by the housing and therefore is not transmitted
through the load cell.
15
3. Biomanufacturing apparatus as claimed in claim 1 or 2, wherein: the load cell is mounted
to the floor of the chamber; the mechanism for rocking is mounted on the load cell; and the tray is
mounted on the mechanism for rocking.
4. Biomanufacturing apparatus as claimed in claim 1,2 or 3, wherein the chamber has a main
chamber region (35) for housing the tkay and the support, and an antechamber region (33)
shallower in depth relative to the door than the main chamber, the antechamber including a panel
(34) to which is mounted the at least one fluid pump device (54,58) such that the fluid handling
25 portion(s) (48,49) of the pun~p(sp) roject beyond the panel into the antechamber.
/i 5. Biomanufacturing apparatus as claimed in claim 4, wherein said at least one connection is
i mounted to the panel, said at least one connection being adapted for removably connecting one or
more of: a gas conduit (106); a pH sensor connection path (108); and a dissolved oxygen sensor
'I ~ 30 connection path 110.
6. Biomanufacturing apparatus as claimed in any one of the preceding claims, wherein thc
tray is slideable relative to, andlor removable from, the tray support.
7. Biomanufacturing apparatus as claimed in claim 6, wherein the door includes a further tray
5 support (120) for supporting a tray on the door when the door is open, the further support being
collapsible to allow the door to close.
8. Biomanufacturing apparatus as claimed in any one of the preceding claims wherein the a
load cell (41), is a a mechanical strain sensor, operable to determine the change in mass supported
I 0 on the tray.
9. Biomanufacturing apparatus as claimed in any one of the preceding claims, further
including a bioreactor heater (42) mounted at the tray for conductive heating of the bioreactor, and
a chamber air heater (53) for convective heating the gaseous atmosphere in the chamber, each
15 heater being controlled by a temperature control.
10. Biomanufacturing apparatus as claimed in any one of the preceding claims wherein said
top and bottom faces are generally flat and include height adjustable feet (26).
20 11. Biomanufacturing apparatus as claimed in any one of the preceding claims, further
including a support (50) external to the housing to one side of the door for supporting consumable
materials, and/or fluid products such as bdgged waste andlor biomolecules.
12. Biomanufacturing apparatus as claimed in claim 5 or as clamed in any one of claims 6 to
25 11 when dependant on claim 5, further including a bioreactor in the form of flexible bag (100)
supported on the tray, said bag including fluid conduits (102,104) passing via said pump heads
(48,49 and paths (106.108.1 10) connected or connectable to said connections (43).
13. Biomanufacturing apparatus as claimed in claim 1112, wherein the bioreactor has a
30 capacity of between approximately 50 millilitres and 2500 millilitres.
14. Biomanufacturing apparatus as claimed in any one of the preceding claims, wherein the
tray or tray support includes a reader (56) for recognising the identity of a bioreactor mounted on
the tray.
5 15. Biomanufacturing apparatus as claimed in any one of the preceding claims, wherein the
chamber, with the door closed, reduces or substantially excludes visible light.
16. Biomanufacturing apparatus as claimed in any one of the preceding claims, wherein the
access door includes a status display (57) viewable from the outside of the door, and/or a window
10 for viewing the chamber.
I
17. A biomanufacturing system including plural stacked biomanufacturing apparatus as
claimed in any one of the preceding claims, in data communication with a cenfral computer (60)
including software operable to monitor the status of andlor control one or more of the plural
15 apparatus.
18.A method for determining mass changes in a flexible bag mounted to a rocking tray inside a
chamber of a cell biomanufacturing apparatus, said method comprising the steps of:
20 a) providing a tray for supporting said flexible bag;
b) providing a rocking mechanism for supporting said tray and for causing roclcing of the
tray; I
c) providing a laod cell supporting the roclcing mechanism to measure the laod on the
rocking mechanism; and either
25 dl) periodically monitoring the static laod on the laod cell to determine changes in the
mass/weight of the flexible bag; or
d2) monitoring the load of the load cell during said roclting motion to determine an arithmetic
mean, median, mode, range or aggregate loading.