WO2020036531A1 - Device and method for preparing water-in-oil and oil-in-water emulsions for immunizations of animals and humans - Google Patents

Device and method for preparing water-in-oil and oil-in-water emulsions for immunizations of animals and humans Download PDF

Info

Publication number
WO2020036531A1
WO2020036531A1 PCT/SE2019/050752 SE2019050752W WO2020036531A1 WO 2020036531 A1 WO2020036531 A1 WO 2020036531A1 SE 2019050752 W SE2019050752 W SE 2019050752W WO 2020036531 A1 WO2020036531 A1 WO 2020036531A1
Authority
WO
WIPO (PCT)
Prior art keywords
emulsion
plunger
sealable opening
syringe
mating arrangement
Prior art date
Application number
PCT/SE2019/050752
Other languages
French (fr)
Inventor
Björn Thomas BÄCKSTRÖM
Original Assignee
Btb Emulsions Ab
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Btb Emulsions Ab filed Critical Btb Emulsions Ab
Priority to EP19849419.7A priority Critical patent/EP3836884A4/en
Publication of WO2020036531A1 publication Critical patent/WO2020036531A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
    • A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
    • A61J1/14—Details; Accessories therefor
    • A61J1/20—Arrangements for transferring or mixing fluids, e.g. from vial to syringe
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M5/00—Devices for bringing media into the body in a subcutaneous, intra-vascular or intramuscular way; Accessories therefor, e.g. filling or cleaning devices, arm-rests
    • A61M5/178—Syringes
    • A61M5/31—Details
    • A61M5/3145—Filters incorporated in syringes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M5/00—Devices for bringing media into the body in a subcutaneous, intra-vascular or intramuscular way; Accessories therefor, e.g. filling or cleaning devices, arm-rests
    • A61M5/178—Syringes
    • A61M5/31—Details
    • A61M5/315—Pistons; Piston-rods; Guiding, blocking or restricting the movement of the rod or piston; Appliances on the rod for facilitating dosing ; Dosing mechanisms
    • A61M5/31511—Piston or piston-rod constructions, e.g. connection of piston with piston-rod
    • A61M5/31515—Connection of piston with piston rod
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
    • B01F23/40—Mixing liquids with liquids; Emulsifying
    • B01F23/41—Emulsifying
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/50—Movable or transportable mixing devices or plants
    • B01F33/501—Movable mixing devices, i.e. readily shifted or displaced from one place to another, e.g. portable during use
    • B01F33/5011—Movable mixing devices, i.e. readily shifted or displaced from one place to another, e.g. portable during use portable during use, e.g. hand-held
    • B01F33/50112—Movable mixing devices, i.e. readily shifted or displaced from one place to another, e.g. portable during use portable during use, e.g. hand-held of the syringe or cartridge type
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
    • B01F35/30—Driving arrangements; Transmissions; Couplings; Brakes
    • B01F35/32—Driving arrangements
    • B01F35/32005—Type of drive
    • B01F35/3202—Hand driven
    • B01F35/32021—Shaking by hand a portable receptacle or stirrer for mixing

Definitions

  • the present invention relates to a device for preparing an emulsion, which is particularly useful when there is a desire to reproduce the quality of an emulsion. Further the invention also relates to a method of preparing an emulsion by use of the device disclosed herein.
  • Freund’s adjuvant is the most common oil used together with antigens dissolved in an aqueous solution.
  • a 2-syringe method, vortexing, or sonication are a few of the methods used for preparing emulsions. These methods are difficult to standardize and there is poor reproducibility between individuals, leading to difficulties in interpreting results from different laboratories. And since the quality of the emulsion is one of the most important features, when using oil-based adjuvants and antigens to generate a satisfactory immune response, reproducibility is important.
  • the most common way to generate a water-in- oil emulsion is to mix the adjuvant and antigen using the, previously mentioned, "syringe” method, allowing the components to pass from one syringe to another.
  • This method has a number of drawbacks such as 1 ) loss of product during preparation and transfer of the ready-made emulsion, 2) time- consuming, due to a long mixing time (>15 minutes), and 3) good emulsions are only obtained in approximately 80% of the emulsions prepared.
  • the present invention discloses such a device and method thereof, which further presents very little loss of product, takes a short time to prepare and is without person-to-person variations.
  • the invention relates to a novel device and use in the preparation of emulsions for vaccines or any other components to be used for immunisation of human or animal to prevent or induce disease, or to produce monoclonal or polyclonal antibodies for diagnostic and therapeutic purposes.
  • the present invention further discloses a device adapted for preparing an emulsion therein, comprising a body with a length, a width, and a cross section, further comprising a first sealable opening for introduction of components to be emulsified, and a mating arrangement, adapted to mate with an external part, wherein the mating arrangement is a protrusion from the body to fit with the opening of an external part, for example a syringe.
  • the protrusion is specifically designed to minimise loss of emulsion upon transfer. In one embodiment, before an external part is mated, a snap-off closure is removed.
  • the present invention discloses a single device for preparing an emulsion, comprising a body with a length, a width, and a cross section, further comprising a first sealable opening, for introduction of components to be emulsified, and a mating arrangement, adapted to mate with an external part, wherein the mating arrangement is a male or female Luer lock fitting.
  • the present invention discloses a device adapted for preparing an emulsion therein, comprising a body with a length, a width, and a cross section, further comprising a first sealable opening, for introduction of components to be emulsified, and a mating arrangement, adapted to mate with an external part, wherein the mating arrangement is made to fit a syringe.
  • the major advantage with the present invention is that the whole emulsion production process takes place in the one and same container, i.e. or in a single container, such as the device according to the present invention.
  • Current standard 2-syringe technique uses two containers to produce emulsions. Creating an emulsion in a single container has the advantages; such as it saves time, is safer, creates reproducible emulsion quality, and diminishes loss of expensive materials that is associated with the 2-syringe technique.
  • a device 1 for preparing an emulsion therein comprising a body 2 with a length 3, a width 4, and a cross section 5, further comprising a first sealable opening 6, for introduction of components to be emulsified, a second sealable opening 15, a mating arrangement 7, wherein the mating arrangement 7 is to mate with a syringe.
  • the sealable opening 15 comprises the mating arrangement 7.
  • the present invention relates to a device 1 for preparing an emulsion therein, comprising a body 2 with a length 3, a width 4, and a cross section 5, further comprising a first sealable opening 6, for introduction of components to be emulsified, a second sealable opening 15 comprising a mating arrangement 7, wherein the mating arrangement 7 is adapted to mate with an external part, such as a syringe.
  • the present invention relates to a device 1 adapted for preparing an emulsion therein, comprising a body 2 with a length 3, a width 4, and a cross section 5, further comprising a first sealable opening 6, for introduction of components to be emulsified, a second sealable opening 15 comprising a mating arrangement 7, wherein the mating arrangement 7 is adapted to mate with a syringe.
  • the second sealable opening 15 may be adapted for introduction of components to be emulsified and/or transfer of an emulsion to an external part, such as a syringe.
  • the first sealable opening 6 comprises a seal selected from the group consisting of cork, plug, stopper, screw-lid, push-down lid, push-down plug, peel-off seal and plunger seal.
  • first sealable opening 6 further comprises threads.
  • mating arrangement 7 and first sealable opening 6 are arranged on opposite sides of the body 2. With opposite sides of the body means the sides of the device extending in the longitudinal direction.
  • the mating arrangement 7 comprises a protrusion 9, preferably the protrusion 9 comprises a snap-off closure 8.
  • the mating arrangement 7 comprises a Luer lock arrangement.
  • the Luer lock may comprise a snap-off closure or another type of seal, such as a cork, a plug, a stopper, a screw cap, a screw-lid, a push down lid, a push-down plug, a peel-off seal or a plunger seal.
  • the mating arrangement 7 comprises hooks 16 and a protrusion 9.
  • the protrusion 9 may comprise a seal, such as a snap-off closure, a cork, a plug, a stopper, a screw cap, a screw-lid, a push down lid, a push-down plug, a peel-off seal or a plunger seal.
  • a seal such as a snap-off closure, a cork, a plug, a stopper, a screw cap, a screw-lid, a push down lid, a push-down plug, a peel-off seal or a plunger seal.
  • the body 2 is elongated, preferably in a tubular shape, preferably made of a material selected from the group consisting of glass, metal or a polymer material, preferably selected from thermoplastics or thermosetting polymers.
  • the device 1 further comprises a flange 1 1 , preferably arranged in close proximity to the first sealable opening 6.
  • the flange 1 1 preferably is a collar, preferably arranged in close proximity to the first sealable opening 6.
  • the device comprises an inner volume, when sealed, of 0.5-500 ml, preferably 1 -500 ml, preferably 1 -100 ml, preferably 1 -50 ml preferably 5-30 ml, preferably 5-15 ml, preferably 5-10 ml.
  • the device comprises an inner volume, when sealed, of 0.5-500 ml, preferably 1 -500 ml, preferably 1 -100 ml, preferably 1 -50 ml, preferably 1 -10 ml, preferably 5-10 ml.
  • the device comprises an inner volume, when sealed, of 1 -10 ml.
  • the device comprises an inner volume, when sealed, of 0.5-2 ml.
  • the inner volume is selected from the group consisting of 500 ml, 400 ml, 300 ml, 250 ml, 200 ml, 100 ml, 50 ml, 30 ml, 15 ml, 10 ml, 5 ml, 1 ml, and 0.5 ml.
  • the present invention relates to a device 1 , comprising a plunger 14 adapted for being inserted through the first sealable opening 6, wherein the plunger comprises a protrusion at the end with an outer thread 17 to fit a plunger seal 19 with an inner thread 18.
  • the device 1 comprises a plunger 20, with an opening at the end 21 , to fit a gas-permeable, liquid non-permeable membrane 22.
  • Another aspect of the present invention relates to a method for creating an emulsion comprising the steps:
  • D 1 centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises the sealed mating arrangement 7;
  • E 1 removing a seal, of the first sealable opening 6 of the device 1 and insert a syringe plunger;
  • the present invention relates to a method for creating an emulsion comprising the steps:
  • the method is further comprising the steps: D 3 ) centrifugation and/or hand shaking of the device 1 , to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises a sealed mating arrangement 7;
  • the present invention relates to a method for creating an emulsion comprising the steps:
  • a 5 filling the device 1 with the components intended to be emulsified; B 5 ) attaching a seal, such as a stop plug at the first sealable opening 6 of the device 1 ; and
  • the present invention relates to a method for creating an emulsion comprising the steps:
  • the device 1 may be filled with components intended to be emulsified through the first sealable opening 6 or the second sealable opening 15 wherein the openings are sealed before the device 1 is shaken to produce an emulsion.
  • the plungers 12, 14 and 20 may be used interchangeably in the device 1 .
  • Yet another aspect of the present invention relates to the use of the device according to the present invention for production of an emulsion.
  • Yet another aspect relates to an emulsion obtained according to the method of the present invention.
  • Figure 1 a Illustrates one embodiment of the present invention from a side view of the device 1 for preparing an emulsion, a body 2 with a length 3, a width 4, one first sealable opening 6, for introduction of components to be emulsified, and threads 10 near the first sealable opening 6 to close the opening with a screw lid or push lid, flanges 1 1 , for easier control during the injection of the emulsion, a mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
  • Figure 1 b Illustrates one embodiment of the present invention from a top view, showing a first sealable opening 6, threads 10, and the flanges 1 1 .
  • Figure 1 c Illustrates one embodiment of the present invention from a cross section view, showing a cross section 5 of the body 2 at point A.
  • Figure 1 d Illustrates one embodiment of the present invention from a bottom view, showing mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
  • Figure 2a Illustrates one embodiment of the present invention from a side view of the device 1 for preparing an emulsion, a body 2 with a length 3, a width 4, one first sealable opening 6, for introduction of components to be emulsified, and threads 10 near the first sealable opening 6 to close the opening with a screw lid, a mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
  • Figure 2b Illustrates one embodiment of the present invention from a top view, showing a first sealable opening 6 and threads 10.
  • Figure 2c Illustrates one embodiment of the present invention from a cross section view, showing a cross section 5 of the body 2 at point A.
  • Figure 2d Illustrates one embodiment of the present invention from a bottom view, showing mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
  • Figure 3a Illustrates one embodiment of the present invention comprising the device 1 for preparing an emulsion, a body 2 with a length 3, a width 4, one first sealable opening 6, for introduction of components to be emulsified, and closed off by a push lid or similar, flanges 1 1 , for easier control during the injection of the emulsion, and a mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
  • Figure 3b Illustrates one embodiment of the present invention from a top view, showing a first sealable opening 6, and flanges 1 1 .
  • Figure 3c Illustrates one embodiment of the present invention from a cross section view, showing a cross section 5 of the body 2 at point A.
  • Figure 3d Illustrates one embodiment of the present invention from a bottom view, showing mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
  • Figure 4a Illustrates part of one embodiment of the present invention from a side view showing a plunger 12 with a plunger seal 13 at one end.
  • Figure 4b Illustrates part of one embodiment of the present invention from a side view showing a plunger 14 comprising outer threads 17 at one end and a plunger seal 19 comprising inner threads 18 throughout the hole of the seal, that will be screwed together before pushing out the emulsion.
  • Figure 4c Illustrates part of one embodiment of the present invention from a side view showing a plunger 20 comprising an end 21 , containing holes to enable air passage when the plunger with a gas-permeable and liquid non- permeable membrane 22 is pushed into the device 1.
  • Figure 4d Illustrates part of one embodiment of the present invention from a bottom view showing an end 21 of a plunger 20.
  • Figure 4e Illustrates part of one embodiment of the present invention showing a gas-permeable and liquid non-permeable membrane 22 that is to be fitted at an end 21 of a plunger 20.
  • Figure 4f Illustrates part of one embodiment of the present invention showing a plunger 20 with an end 21 fitted with a gas-permeable and liquid non-permeable membrane 22.
  • FIG. 5 Illustrates results of a comparative study performed by an independent CRO for an emulsion prepared by the method of the present invention ( ⁇ ) and an emulsion prepared by the commonly used“2-syringe method” (O).
  • EAE Experimental Autoimmune Encephalomyelitis
  • MOG Myelin protein
  • CFA complete Freund’s Adjuvant
  • the invention relates to a novel single device and method to prepare water-in oil and oil-in water emulsions, to induce immune responses in animals and humans.
  • the combination of the device and the method will ensure rapid, consistent, and almost waste-free production of high quality emulsions.
  • the invention is interrelated with: 1 ) induction of disease in autoimmune animal models, 2) testing potential immunotherapies of cancer treatments, 3) preparation of vaccines for use of immunization of humans and animals, and 4) immunization of animals for production of monoclonal and polyclonal antibodies for diagnostic and therapeutic purposes.
  • an "adjuvant” an immune enhancing substance
  • an antigen a virus, bacteria, protein, peptides or haptens.
  • the most commonly used oil adjuvant to create a water-in-oil emulsion is Freund’s adjuvant for immunizing animals.
  • Freund a Perfect Oil & Water Emulsion Receptacle
  • the main element of the invention is a single device 1 , that is to be used together with a shaking machine to prepare oil and water emulsions (water-in- oil and oil-in-water).
  • emulsion may be interpreted as a dispersion of two liquids that are normally immiscible, whereas the term suspension may be interpreted as a heterogeneous mixture that contains solid particles.
  • the device of the present invention may also be used to prepared suspensions.
  • the device 1 looks like a tube with one opening at one end 6, and a mating arrangement
  • the mating arrangement 7 may be a snap-off closure 8.
  • the mating arrangement may be fitted with other types of seals, such as cork, plug, stopper, screw cap, screw-lid, push-down lid, push-down plug, a peel-off seal, or with a plunger seal.
  • first adjuvants and antigens will be added by pipetting or other means, and then the first sealable opening 6, will be closed with a seal which may be a cork, plug, stopper, screw cap, screw-lid, push-down lid, push-down plug, a peel-off seal or with a plunger-seal.
  • the device 1 will then be shaken in a high speed "shaking machine" at a speed, for a time, depending on which machine is used to produce a perfect emulsion. In a perfect emulsion there is no distinguishable phase separation in the emulsion. To remove any air trapped in the emulsion, the device 1 , will be centrifuged for a short time at a low g-force. Finally, the emulsion will be transferred to a syringe.
  • the seal of the of the first sealable opening 6 of the device 1 will be removed and a plunger 12 with a plunger seal 13 attached will be used to push the emulsion into a syringe, or a plunger 14, with an outer thread 17 at one end, to be attached to a plunger seal 19 with an inner thread 18 with a short turn so that air can go through the threads 17 and 18, then it will be pushed in to the top of the emulsion to eliminate air trapped, then the plunger is turned so that it closes off the hole in the plunger seal 19.
  • plunger means that any plunger or plunger rod available and suitable for this purpose are included in the present invention.
  • the device 1 for preparing an emulsion, comprising a body 2 with a length 3, a width 4, and a cross section 5. Further, the body 2 comprise one first sealable opening 6, for introduction of components to be emulsified, and one opening comprising a snap-off closure 8.
  • the protrusion 9 left after removal of the snap-off closure 8 together with the hooks 16 is designed to fit the back opening of a syringe.
  • One purpose of the invention is to facilitate transfer of a created emulsion into a syringe without any trapped air.
  • the body 2 of the device 1 is elongated. In another embodiment the body 2 is a tubular shape. In yet another embodiment the body 2 is a cylindrical shape, but also other geometrical shapes are thinkable.
  • the device 1 according to the present invention should not be seen as limited by a certain geometrical shape. Further, the device 1 , may be of various materials. In one embodiment the device 1 , is made of glass, metal and/or a polymer material. In one embodiment the polymer material is selected from thermoplastics or thermosetting polymers.
  • the present invention may be comprising, acrylic (polymethyl methacrylate, PMMA), Polycarbonate (PC), Polyvinyl chloride (PVC), Polypropylene (PP), Polyethylene terephthalate (PET / APET), PETG, Acrylonitrile Butadiene Styrene (ABS) and fluorinated polymers, such as fluorinated ethylene-propylene (FEP), polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF), perfluoroalkoxy alkane (PFA) and/or any combinations thereof.
  • FEP fluorinated ethylene-propylene
  • PTFE polytetrafluoroethylene
  • PVDF polyvinylidene fluoride
  • PFA perfluoroalkoxy alkane
  • the device 1 comprising an opening 6, for introduction of components to be emulsified.
  • the opening 6 will then be closed with a seal.
  • seals may be seals selected from the group consisting of screw lid, push lid, plug, thermoplastic plunger stopper, plunger seal, peel-off seal, cork or other devices that may seal one end of the device.
  • a plunger 12 with a plunger seal 13 or a plunger 14 with an outer thread 17, together with a plunger seal 19 with an inner thread 18, may be inserted into the first sealable opening 6 (the top) of the device 1 .
  • the seal is a plunger head. In one embodiment the seal is adapted to mate with a plunger tip. In another embodiment the plunger comprises inner threads and the plunger tip outer threads. In yet another embodiment, the plunger 20 is fitted with a gas-permeable and liquid non- permeable membrane 22, which will be mounted into an opening at the end 21 . The end 21 has holes through the base where air can escape when the plunger is pushed into the syringe with emulsions.
  • the plunger-tip comprises a recess optionally comprising an opening with a narrower entrance portion to which a plunger with a protrusion at the end that can be fitted.
  • a common seal is a screw lid.
  • the body 2 of the device 1 of the present invention comprise threads 10. These threads 10 may be arranged in close proximity to the first sealable opening 6 of the device 1 where the components to be emulsified, are to be introduced. Further, the threads may be arranged to mate with the threads of a screw lid. Various angles of the threads are thinkable.
  • the top of the device comprises an inner thread and the screw-lid comprise an outer thread. The outer thread of the screw lid is to be fitted with the inner thread of the device.
  • mating arrangement 7 of the device 1 comprising hooks 16 and a protrusion 9, is adapted to fit the back end of a syringe.
  • the mating arrangement 7 fits an adaptor, for example a Luer Lock arrangement. This may be helpful for fitting syringes that are of larger sizes. Since one of the purposes of the present invention is to facilitate transfer of created emulsion, in one embodiment, the mating arrangement 7 comprise threads at the end, preferably outer threads.
  • the present invention comprises a female-to-female, male-to-male or male-to-female adaptor.
  • this embodiment may result in some additional loss of products, compared to the embodiments not using this adaptor.
  • One application for emulsions may be, as mentioned, to be administered by a syringe.
  • the mating arrangement 7 is adapted to mate with a syringe.
  • the device 1 may be of various sizes and is not to be seen as limited by a specific size.
  • the length 3 is larger than the width 4.
  • the length 3 is selected from the group consisting of 10-500 mm, 10-300 mm, 10-200 mm, 10- 150 mm, 10-100 mm, 10-50 mm, and/or the width 4 is selected from the group consisting of 5-100 mm or 5-50 mm.
  • the relationship between the length 3 and the width 4 (lengtlrwidth) is at least 2:1 , at least 3:1 , at least 4:1 , at least 5:1 , at least 6:1 , at least 7:1 , at least 8:1 , at least 9:1 , at least 10:1 .
  • the relationship between the length 3 and the width 4 is at most 15:1 .
  • the device 1 may comprise various geometrical shapes.
  • the cross section 5, seen across the length 3 of the body 2 comprise a geometrical shape such as circular, oval, square or square with rounded corners, triangular, triangular with rounded corners.
  • the cross section 5 comprises a geometrical shape with corners such as at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten corners.
  • the cross section 5 comprises at most 20 corners.
  • a flange 1 1 may be arranged in close proximity to the first sealable opening 6.
  • the flange 1 1 may be a collar or a skirt.
  • the flange 1 1 at least partially encloses the body 2 along part of the length 3.
  • the flange 1 1 may be the shape of a ring, a collar, wings or similar to those on a syringe. Other shapes are thinkable and the purpose of the flange 1 1 is to stop the device from slipping through the finger while pushing the plunger into the device.
  • the flange 1 1 is in one embodiment 1 -50 mm larger in radius than the cross section of the device 1 .
  • the device 1 may have different inner volumes.
  • the device 1 comprises an inner volume, when sealed, of preferably 0.5-100 ml, preferably 1 -50 ml.
  • the device 1 comprises an inner volume, selected from the group consisting of 0.5-5 ml, 1 -10 ml, 5-15 ml, 5-30 ml, 10-50 ml, 10-100 ml, and 50-500 ml.
  • the present invention is a method for creating an emulsion comprising the steps:
  • the method further comprises a centrifugation step after shaking in step C 8 ).
  • the method further comprises removing the screw lid or push lid and inserting a plunger at the top, or inserting a plunger at the top containing an outer thread to fit the plunger-tip in the device with an inner thread, or inserting a plunger with a protrusion at the end to fit the plunger seal in the device comprising a recess opening with a narrower entrance portion, or inserting a plunger fitted with a gas-permeable, liquid non-permeable membrane at its end, and then removing the snap-off closure at the other end and attach a syringe.
  • step B 8 ) of the present invention further comprises closing the first sealable opening 6 of the device 1 with a seal.
  • the device 1 looks like a tube with one opening at one end, a first sealable opening 6, and a female Luer Lock adapter at the other end, a second sealable opening 15.
  • a male Luer Lock Cap will be attached to the female Luer Lock fitting, adjuvants and antigens will then be added by pipetting or other means, and finally the first sealable opening 6, is closed with a seal wherein the seal may be a screw cap, screw-lid, push-down lid, push-down plug or with a plunger seal and a screw cap.
  • the device 1 will then be shaken in a high speed "shaking machine" at a speed, for a time, depending on the machine used, to produce a perfect emulsion.
  • the time for preparing an emulsion of high quality with the method of the present invention is preferably 0.5-12 min, preferably 1 -10 min, preferably, 2-8 min, preferably 5-7 min. In a perfect emulsion there is no distinguishable phase separation in the emulsion. To remove any air trapped in the emulsion, the device 1 , will be centrifuged for a short time at a low en force.
  • any syringe of either male Luer Lock or Luer Slip type will be attached to the device 1 , screw lid removed and a plunger, attached to push the emulsion into the syringe, or a plunger, to be attached to the plunger seal 19 with a short turn so that air can go through the threads 18, then it is pushed in to a close proximity of the emulsion, then the plunger is turned so that it closes off the hole in plunger seal, and then the emulsion is pushed out into the attached syringe.
  • the device 1 for preparing an emulsion, comprising a body 2 with a length 3, a width 4, and a cross section 5. Further, the body 2 comprise a first sealable opening 6, for introduction of components to be emulsified, and a second sealable opening 15 comprising a female Luer Lock fitting.
  • the female Luer lock fitting may be designed to fit a male Luer Lock fitting and/or a Slip fitting of a syringe.
  • One purpose of the invention is to facilitate transfer of a created emulsion into a syringe, preferably under aseptic conditions.
  • the body 2 of the device 1 is elongated.
  • the body 2 is a tubular shape.
  • the body 2 is a cylindrical shape, but also other geometrical shapes are thinkable.
  • the device 1 according to the present invention should not be seen as limited by a certain geometrical shape.
  • the device 1 further comprises a seal, arranged in close proximity to the first sealable opening 6, for introduction of components to be emulsified.
  • the first sealable opening 6 is a sealable end.
  • the sealable end 6 may be sealed with a seal.
  • seals are thinkable, examples of seals may be seals selected from the group consisting of screw lid, push lid, plug, stopper, plunger-tip, cork or other devices that may seal one end of the device. In the present application the wording seal and close are used interchangeably.
  • a plunger seal 13 and/or a plunger seal 19, with an inner thread 18, through the hole of plunger seal may be inserted into the first opening 6 of the device 1 .
  • the seal may further comprise a plunger 14, with outer threads 17 at the end that can be fitted with the plunger seal 19 with inner threads 18.
  • the seal is a plunger head.
  • the plunger seal comprises a recess optionally comprising an opening with a narrower entrance portion to which a plunger with a protrusion at the end that can be fitted.
  • a common seal for the first sealable opening 6 is a screw lid.
  • the body 2 of the device 1 of the present invention comprise threads 10. These threads may be arranged in close proximity to the first sealable opening 6 of the device 1 where the components to be emulsified, are to be introduced. Further, the threads may be arranged to mate with the threads of a screw lid. Various angles of the threads are thinkable.
  • the top of the device comprises an inner thread and the screw-lid comprise an outer thread. The outer thread of the screw lid is to be fitted with the inner thread of the device.
  • mating arrangement 7 of the device is adapted to fit male Luer lock fittings preferably of syringes.
  • the mating arrangement 7 fits an adaptor. This may be helpful for fitting syringes that lack Luer lock. Since one of the purposes of the present invention is to facilitate transfer of created emulsion, in one embodiment, the mating arrangement 7 comprise threads at the end, preferably outer threads.
  • the present invention comprises a female-to-female, male-to-male or male-to-female adaptor.
  • this embodiment may result in some additional loss of products compared to not using this adaptor.
  • One application for emulsions may be, as mentioned, to be administered by a syringe.
  • the mating arrangement 7 is adapted to mate with a syringe.
  • the device 1 may be of various sizes and is not to be seen as limited by a specific size.
  • the length 3 is larger than the width 4.
  • the length 3 is selected from the group consisting of 10-500 mm, 10-300 mm, 10-200 mm, 10- 150 mm, 10-100 mm, 10-50 mm, and/or the width 4 is selected from the group consisting of 5-100 mm or 5-50 mm.
  • the relationship between the length 3 and the width 4 (lengtlrwidth) is at least 2:1 , at least 3:1 , at least 4:1 , at least 5:1 , at least 6:1 , at least 7:1 , at least 8:1 , at least 9:1 , at least 10:1 .
  • the relationship between the length 3 and the width 4 is at most 15:1 .
  • the device 1 may comprise various geometrical shapes.
  • the cross section 5, seen across the length 3 of the body 2 comprise a geometrical shape such as circular, oval, square or square with rounded corners, triangular, triangular with rounded corners.
  • the cross section 5 comprises a geometrical shape with corners such as at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten corners.
  • the cross section 5 comprises at most 20 corners.
  • a flange 1 1 may be arranged in close proximity to the first sealable opening 6.
  • the flange 1 1 may be a collar or a skirt.
  • the flange 1 1 at least partially encloses the body 2 along part of the length 3.
  • the collar may be the shape of a ring, a flange, wings or similar to those on a syringe. Other shapes are thinkable and the purpose of the flange 1 1 is to stop the device from slipping through the finger while pushing the plunger into the device.
  • the flange 1 1 is in one embodiment 1 -50 mm larger in radius than the cross section of the device 1 .
  • the device 1 may have different inner volumes.
  • the device 1 comprises an inner volume, when sealed, of preferably 0.5-100 ml, preferably 1 -50 ml.
  • the device 1 comprises an inner volume, selected from the group consisting of 0.5-5 ml, 1 -10 ml, 5-15 ml, 5-30 ml, 10-50 ml, 10-100 ml, and 50-500 ml.
  • the present invention is a method for creating an emulsion comprising the steps:
  • a 9 attaching a stop plug to the mating arrangement 7 near the second sealable opening 15 of the device 1 according to the present invention
  • the method further comprises a centrifugation step after shaking in step D 9 ).
  • the mating arrangement 7 may be a female Luer Lock fitting.
  • the method further comprises removing the screw lid or push lid and inserting a plunger at the top, or inserting a plunger at the top containing an outer thread to fit the plunger seal in the device with an inner thread, or inserting a plunger with a protrusion at the end to fit the plunger seal in the device comprising a recess opening with a narrower entrance portion, or inserting a plunger fitted with a gas-permeable, liquid non-permeable membrane at its end, and then removing the stop plug at the other end and attach a male Luer lock or Luer slip syringe.
  • Another aspect of the present invention relates to a method for creating an emulsion comprising the steps:
  • the method is further comprising the steps: D 10 ) centrifugation and/or hand shaking of the device 1 to force the emulsion towards a sealed second sealable opening 15 of the device 1 , which second sealable opening 15 comprise the mating arrangement 7;
  • Another aspect of the present invention relates to a method for creating an emulsion comprising the steps:
  • a 11 attaching a stop plug to the female Luer Lock fitting at the second sealable opening of the device 1 according to the present invention
  • the method further comprises a step to use a plunger containing a protrusion at the end with an outer thread to push in the plunger seal with an inner thread to close proximity of the surface of the emulsion without screwing in the plunger to let the air out, and then fasten the plunger to the plunger seal before forcing the emulsion through the second sealable opening and into an attached syringe.
  • the method further comprise pushing out the emulsion into the syringe by pressing in the plunger at the top and stopping the device to move by holding onto the protrusion.
  • Another aspect of the present invention relates to a kit of parts comprising the device according to the present invention, and an instruction pamphlet comprising the method according to what is disclosed herein, regarding the present invention.
  • the present invention relates to use of the device according to what is disclosed herein for production of an emulsion.
  • the invention relates to an emulsion obtained according to the method of the present invention, described herein.
  • the present invention is suitable for preparing liposomes.
  • the device and method described here have a number of advantages.
  • FIG. 5 shows results of a comparative study for an emulsion prepared by the method of the present invention and a control emulsion, by using the Experimental Autoimmune Encephalomyelitis (EAE) model, which is an animal disease model used to study the human disease Multiple Sclerosis.
  • EAE Experimental Autoimmune Encephalomyelitis
  • Preparation of a high-quality water-in-oil emulsion containing Myelin protein is necessary to induce disease in research animals.
  • Animals were injected with an emulsion consisting of Myelin protein (MOG) mixed with complete Freund’s Adjuvant (CFA) prepared using two different methods.
  • Animals were scored daily for disease using the grades 0-8, depending on the degree of paralysis of each animal. Each group consisted of 10 animals. Shown are the mean disease scores ⁇ SEM (standard error of mean
  • the present invention provides a homogenous emulsion that is independent on personal differences.
  • the common 2-syringe method varies from person to person and can be of very different quality, depending on finger strength, time and speed used, temperature in the laboratory, just mentioning a few parameters.
  • When prepared using the 2-syringe method a large proportion of the emulsion is lost in the parts used.
  • the device according to the present invention may comprise different standard components, also having varied design, and being manufactured in various thinkable materials which are not explicitly mentioned. Moreover, the design of the device may vary, and the present invention, as formulated in the present claim set, should be seen as embodying different forms and materials of the device.

Abstract

The present invention relates to a device for preparing an emulsion therein, comprising a body with a length, a width, and a cross section, further comprising one first sealable opening, for introduction of components to be emulsified, and a mating arrangement, adapted to mate with an external part, wherein the mating arrangement consists of hooks and a protrusion. The present invention also relates to a method and use of a device for preparing an emulsion.

Description

DEVICE AND METHOD FOR PREPARING WATER-IN-OIL AND OIL-IN-
WATER EMULSIONS FOR IMMUNIZATIONS OF ANIMALS AND HUMANS
Field of the invention
The present invention relates to a device for preparing an emulsion, which is particularly useful when there is a desire to reproduce the quality of an emulsion. Further the invention also relates to a method of preparing an emulsion by use of the device disclosed herein.
Technical Background
When working with potential biopharmaceuticals to treat autoimmune diseases and cancers with immunotherapy, a number of pre-clinical animal models exist. To induce disease in for example the EAE (experimental autoimmune encephalomyelitis) model for human MS (Multiple Sclerosis), CIA (collagen-induced arthritis) model for RA (Rheumatoid Arthritis), and MG (myasthenia gravis), a perfect water-in-oil emulsion is necessary. In addition, to induce a strong immune response to eradicate cancerous cells in the body, also here a perfect water-in-oil emulsion is necessary. To prepare these water- in-oil emulsions for pre-clinical studies, Freund’s adjuvant is the most common oil used together with antigens dissolved in an aqueous solution. Until today, a 2-syringe method, vortexing, or sonication are a few of the methods used for preparing emulsions. These methods are difficult to standardize and there is poor reproducibility between individuals, leading to difficulties in interpreting results from different laboratories. And since the quality of the emulsion is one of the most important features, when using oil-based adjuvants and antigens to generate a satisfactory immune response, reproducibility is important.
Among these methods, the most common way to generate a water-in- oil emulsion is to mix the adjuvant and antigen using the, previously mentioned, "syringe" method, allowing the components to pass from one syringe to another. This method has a number of drawbacks such as 1 ) loss of product during preparation and transfer of the ready-made emulsion, 2) time- consuming, due to a long mixing time (>15 minutes), and 3) good emulsions are only obtained in approximately 80% of the emulsions prepared.
There is therefore a need for a device and method, which eliminates person-to-person differences along all the difficulties presented above. The present invention discloses such a device and method thereof, which further presents very little loss of product, takes a short time to prepare and is without person-to-person variations.
Summary of the invention
The invention relates to a novel device and use in the preparation of emulsions for vaccines or any other components to be used for immunisation of human or animal to prevent or induce disease, or to produce monoclonal or polyclonal antibodies for diagnostic and therapeutic purposes.
The present invention further discloses a device adapted for preparing an emulsion therein, comprising a body with a length, a width, and a cross section, further comprising a first sealable opening for introduction of components to be emulsified, and a mating arrangement, adapted to mate with an external part, wherein the mating arrangement is a protrusion from the body to fit with the opening of an external part, for example a syringe. The protrusion is specifically designed to minimise loss of emulsion upon transfer. In one embodiment, before an external part is mated, a snap-off closure is removed. In one embodiment the present invention discloses a single device for preparing an emulsion, comprising a body with a length, a width, and a cross section, further comprising a first sealable opening, for introduction of components to be emulsified, and a mating arrangement, adapted to mate with an external part, wherein the mating arrangement is a male or female Luer lock fitting. In another embodiment the present invention discloses a device adapted for preparing an emulsion therein, comprising a body with a length, a width, and a cross section, further comprising a first sealable opening, for introduction of components to be emulsified, and a mating arrangement, adapted to mate with an external part, wherein the mating arrangement is made to fit a syringe. The major advantage with the present invention is that the whole emulsion production process takes place in the one and same container, i.e. or in a single container, such as the device according to the present invention. Current standard 2-syringe technique uses two containers to produce emulsions. Creating an emulsion in a single container has the advantages; such as it saves time, is safer, creates reproducible emulsion quality, and diminishes loss of expensive materials that is associated with the 2-syringe technique.
In one aspect of the present invention relates to a device 1 for preparing an emulsion therein, comprising a body 2 with a length 3, a width 4, and a cross section 5, further comprising a first sealable opening 6, for introduction of components to be emulsified, a second sealable opening 15, a mating arrangement 7, wherein the mating arrangement 7 is to mate with a syringe.
In one embodiment the sealable opening 15 comprises the mating arrangement 7.
In one embodiment the present invention relates to a device 1 for preparing an emulsion therein, comprising a body 2 with a length 3, a width 4, and a cross section 5, further comprising a first sealable opening 6, for introduction of components to be emulsified, a second sealable opening 15 comprising a mating arrangement 7, wherein the mating arrangement 7 is adapted to mate with an external part, such as a syringe. In one embodiment the present invention relates to a device 1 adapted for preparing an emulsion therein, comprising a body 2 with a length 3, a width 4, and a cross section 5, further comprising a first sealable opening 6, for introduction of components to be emulsified, a second sealable opening 15 comprising a mating arrangement 7, wherein the mating arrangement 7 is adapted to mate with a syringe.
Optionally, the second sealable opening 15 may be adapted for introduction of components to be emulsified and/or transfer of an emulsion to an external part, such as a syringe.
In one embodiment the first sealable opening 6 comprises a seal selected from the group consisting of cork, plug, stopper, screw-lid, push-down lid, push-down plug, peel-off seal and plunger seal.
In another embodiment the first sealable opening 6 further comprises threads. In another embodiment the mating arrangement 7 and first sealable opening 6 are arranged on opposite sides of the body 2. With opposite sides of the body means the sides of the device extending in the longitudinal direction.
In another embodiment the mating arrangement 7 comprises a protrusion 9, preferably the protrusion 9 comprises a snap-off closure 8.
In one embodiment the mating arrangement 7 comprises a Luer lock arrangement. The Luer lock may comprise a snap-off closure or another type of seal, such as a cork, a plug, a stopper, a screw cap, a screw-lid, a push down lid, a push-down plug, a peel-off seal or a plunger seal.
In yet another embodiment the mating arrangement 7 comprises hooks 16 and a protrusion 9.
In another embodiment the protrusion 9 may comprise a seal, such as a snap-off closure, a cork, a plug, a stopper, a screw cap, a screw-lid, a push down lid, a push-down plug, a peel-off seal or a plunger seal.
In another embodiment the body 2 is elongated, preferably in a tubular shape, preferably made of a material selected from the group consisting of glass, metal or a polymer material, preferably selected from thermoplastics or thermosetting polymers.
In another embodiment the device 1 further comprises a flange 1 1 , preferably arranged in close proximity to the first sealable opening 6.
In one embodiment the flange 1 1 , preferably is a collar, preferably arranged in close proximity to the first sealable opening 6.
In one embodiment the device comprises an inner volume, when sealed, of 0.5-500 ml, preferably 1 -500 ml, preferably 1 -100 ml, preferably 1 -50 ml preferably 5-30 ml, preferably 5-15 ml, preferably 5-10 ml.
In one embodiment the device comprises an inner volume, when sealed, of 0.5-500 ml, preferably 1 -500 ml, preferably 1 -100 ml, preferably 1 -50 ml, preferably 1 -10 ml, preferably 5-10 ml.
In one embodiment the device comprises an inner volume, when sealed, of 1 -10 ml.
In one embodiment the device comprises an inner volume, when sealed, of 0.5-2 ml. In one embodiment the inner volume is selected from the group consisting of 500 ml, 400 ml, 300 ml, 250 ml, 200 ml, 100 ml, 50 ml, 30 ml, 15 ml, 10 ml, 5 ml, 1 ml, and 0.5 ml.
In one embodiment the present invention relates to a device 1 , comprising a plunger 14 adapted for being inserted through the first sealable opening 6, wherein the plunger comprises a protrusion at the end with an outer thread 17 to fit a plunger seal 19 with an inner thread 18.
In a further embodiment the device 1 according to the present invention comprises a plunger 20, with an opening at the end 21 , to fit a gas-permeable, liquid non-permeable membrane 22.
Another aspect of the present invention relates to a method for creating an emulsion comprising the steps:
A) filling the device 1 according to the present invention with the components intended to be emulsified;
B) closing the first sealable opening 6 of the device 1 ; and
C) shaking the closed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 comprising the mating arrangement 7;
E) connecting a syringe to the mating arrangement 7;
F) forcing the emulsion from the device 1 , through the mating arrangement 7, and into the connected syringe.
In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A1) filling the device 1 with the components intended to be emulsified;
B1) sealing the first sealable opening 6 of the device 1 ; and
C1) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D1) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises the sealed mating arrangement 7; E1) removing a seal, of the first sealable opening 6 of the device 1 and insert a syringe plunger;
F1) pushing the syringe plunger into the device 1 until it reaches the emulsion;
G1) connecting a syringe to the mating arrangement 7; and
H1) forcing the emulsion through the second sealable opening 15 and into the connected syringe.
In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A2) filling the device 1 with the components intended to be emulsified;
B2) closing the first sealable opening 6 of the device 1 ; and
C2) shaking the closed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D2) centrifugation and/or hard hand shaking of the device 1 , to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises the sealed mating arrangement 7;
E2) removing a seal, of the first sealable opening 6 of the device 1 and insert a syringe plunger;
F2) pushing the syringe plunger into the device 1 until it reaches the emulsion;
G2) removing a snap-off closure 8 of the mating arrangement 7, and connecting a syringe, to the mating arrangement 7; and
FI2) pushing the syringe plunger into the device 1 to force the emulsion through the second sealable opening 15 and into the connected syringe. In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A3) filling the device 1 with the components intended to be emulsified through the first sealable opening 6;
B3) closing the first sealable opening 6 of the device 1 with a seal; and C3) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps: D3) centrifugation and/or hand shaking of the device 1 , to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises a sealed mating arrangement 7;
E3) removing the seal, of the first sealable opening 6 of the device 1 and insert a syringe plunger and pushing it in until it reaches the emulsion;
F3) removing a snap-off closure 8 of the mating arrangement 7, and connecting the back opening of a syringe to the mating arrangement 7 and locking it into the hooks 16, part of the mating arrangement 7; and
G3) pushing the syringe plunger into the device 1 to force the emulsion through the second sealable opening 15 and into the connected syringe. In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A4) filling the device 1 with the components intended to be emulsified;
B4) closing the first sealable opening 6 of the device 1 with a screw lid;
C4) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D4) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises a sealed mating arrangement 7;
E4) removing the seal, of the first sealable opening 6 of the device 1 and insert a syringe plunger 12 with a plunger seal 13 and pushing it in until it reaches the emulsion;
F4) removing a seal 8 of the mating arrangement 7;
G4) connecting an adaptor that will make it possible to connect a Luer lock syringe to the mating arrangement 7; and
FI4) pushing the syringe plunger 12 with the plunger seal 13 further into the device 1 to force the emulsion through the second sealable opening 15 comprises the mating arrangement 7, and into the connected syringe.
In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A5) filling the device 1 with the components intended to be emulsified; B5) attaching a seal, such as a stop plug at the first sealable opening 6 of the device 1 ; and
C5) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D5) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises a sealed mating arrangement 7;
E5) removing the seal, of the first sealable opening 6 of the device 1 and insert a syringe plunger 12 with a plunger seal 13 and pushing it in until it reaches the emulsion;
F5) removing a seal 8 of the mating arrangement 7;
G5) connecting a male or female Luer Lock or Luer Slip syringe to the mating arrangement 7 or connecting an adaptor that will make it possible to connect a Luer lock syringe to the mating arrangement 7;
H5) pushing the syringe plunger into the device 1 to force the emulsion through the mating arrangement 7 of the second sealable opening 15 and into the connected syringe.
In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A6) filling the device 1 with the components intended to be emulsified;
B6) closing the first sealable opening 6 of the device 1 with a push lid; and
C6) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D6) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises a sealed mating arrangement 7;
E6) removing the seal, of the first sealable opening 6 of the device 1 and insert a syringe plunger 12 with a plunger seal 13 and pushing it in until it reaches the emulsion;
F6) removing a seal 8 of the mating arrangement 7; G6) and connecting the back opening of a syringe to the mating arrangement 7 and locking it in close proximity of a protrusion 9 and hooks 16, part of the mating arrangement 7; and
H6) pushing the syringe plunger 12 the plunger seal 13 further into the device 1 to force the emulsion through the second sealable opening 15 and into the connected syringe.
In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A7) filling the device 1 with the components intended to be emulsified;
B7) closing the first sealable opening 6 of the device 1 with a screw lid or push lid; and
C7) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D7) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises a sealed mating arrangement 7;
E7) removing the screw lid or push lid, of the first sealable opening 6 of the device 1.
F7) inserting a plunger seal 19 with an inner thread 18 at the top device 1 ;
G7) attaching a plunger 14 with an outer thread at the end 17 to fit the plunger seal 19, with a short turn so that air can go through the threads 17 and 18;
H7) pushing the plunger 14 in until it reaches the emulsion to eliminate air trapped;
I7) turning the plunger 14 so that it closes off the hole in the seal 19;
J7) removing a seal 8 of the mating arrangement 7, and connecting the back opening of a syringe to the mating arrangement 7 and locking it into the hooks 16, part of the mating arrangement 7;
K7) pushing the syringe plunger further into the device 1 to force the emulsion through the second sealable opening 15 and into the connected syringe. In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A12) filling the device 1 with the components intended to be emulsified;
B12) closing the first sealable opening 6 of the device 1 with a screw lid;
C12) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D12) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises a sealed mating arrangement 7;
E12) removing the seal, of the first sealable opening 6 of the device 1 and insert a syringe plunger 20 fitted with a gas-permeable, liquid non- permeable membrane 22, pushing the plunger 20 in until it reaches the emulsion to eliminate air trapped;
F12) removing a seal 8 of the mating arrangement 7;
G12) connecting an adaptor that will make it possible to connect a Luer lock syringe to the mating arrangement 7; and
H12) pushing the syringe plunger 20 with the membrane 22 further into the device 1 to force the emulsion through the second sealable opening 15 and into the connected syringe.
In one embodiment the present invention relates to a method for creating an emulsion comprising the steps:
A13) filling the device 1 with the components intended to be emulsified through the second sealable opening 15, which comprises the mating arrangement 7;
B13) closing the first sealable opening 6 and the second sealable opening 15 comprising the mating arrangement 7 with a seal; and
C13) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
D13) centrifugation and/or hand shaking of the device 1 , to force the emulsion towards the second sealable opening 15, which comprises the sealed mating arrangement 7; E13) insert a syringe plunger into the first sealable opening 6 of the device 1 and pushing the syringe plunger in until it reaches the emulsion to eliminate air trapped;
F13) removing the seal of the mating arrangement 7, and connecting a syringe to the mating arrangement 7 and locking it into the mating arrangement 7; and
G13) pushing the syringe plunger into the device 1 to force the emulsion through the second sealable opening 15 and into the connected syringe. In yet another embodiment the device 1 may be filled with components intended to be emulsified through the first sealable opening 6 or the second sealable opening 15 wherein the openings are sealed before the device 1 is shaken to produce an emulsion. The plungers 12, 14 and 20 may be used interchangeably in the device 1 .
Yet another aspect of the present invention relates to the use of the device according to the present invention for production of an emulsion.
And yet another aspect relates to an emulsion obtained according to the method of the present invention.
Short description of the drawings
Figure 1 a. Illustrates one embodiment of the present invention from a side view of the device 1 for preparing an emulsion, a body 2 with a length 3, a width 4, one first sealable opening 6, for introduction of components to be emulsified, and threads 10 near the first sealable opening 6 to close the opening with a screw lid or push lid, flanges 1 1 , for easier control during the injection of the emulsion, a mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
Figure 1 b. Illustrates one embodiment of the present invention from a top view, showing a first sealable opening 6, threads 10, and the flanges 1 1 .
Figure 1 c. Illustrates one embodiment of the present invention from a cross section view, showing a cross section 5 of the body 2 at point A. Figure 1 d. Illustrates one embodiment of the present invention from a bottom view, showing mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
Figure 2a. Illustrates one embodiment of the present invention from a side view of the device 1 for preparing an emulsion, a body 2 with a length 3, a width 4, one first sealable opening 6, for introduction of components to be emulsified, and threads 10 near the first sealable opening 6 to close the opening with a screw lid, a mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
Figure 2b. Illustrates one embodiment of the present invention from a top view, showing a first sealable opening 6 and threads 10.
Figure 2c. Illustrates one embodiment of the present invention from a cross section view, showing a cross section 5 of the body 2 at point A.
Figure 2d. Illustrates one embodiment of the present invention from a bottom view, showing mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
Figure 3a. Illustrates one embodiment of the present invention comprising the device 1 for preparing an emulsion, a body 2 with a length 3, a width 4, one first sealable opening 6, for introduction of components to be emulsified, and closed off by a push lid or similar, flanges 1 1 , for easier control during the injection of the emulsion, and a mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
Figure 3b. Illustrates one embodiment of the present invention from a top view, showing a first sealable opening 6, and flanges 1 1 .
Figure 3c. Illustrates one embodiment of the present invention from a cross section view, showing a cross section 5 of the body 2 at point A.
Figure 3d. Illustrates one embodiment of the present invention from a bottom view, showing mating arrangement 7 comprising hooks 16 and a protrusion 9, and a snap-off closure 8.
Figure 4a. Illustrates part of one embodiment of the present invention from a side view showing a plunger 12 with a plunger seal 13 at one end. Figure 4b. Illustrates part of one embodiment of the present invention from a side view showing a plunger 14 comprising outer threads 17 at one end and a plunger seal 19 comprising inner threads 18 throughout the hole of the seal, that will be screwed together before pushing out the emulsion.
Figure 4c. Illustrates part of one embodiment of the present invention from a side view showing a plunger 20 comprising an end 21 , containing holes to enable air passage when the plunger with a gas-permeable and liquid non- permeable membrane 22 is pushed into the device 1.
Figure 4d. Illustrates part of one embodiment of the present invention from a bottom view showing an end 21 of a plunger 20.
Figure 4e. Illustrates part of one embodiment of the present invention showing a gas-permeable and liquid non-permeable membrane 22 that is to be fitted at an end 21 of a plunger 20.
Figure 4f. Illustrates part of one embodiment of the present invention showing a plunger 20 with an end 21 fitted with a gas-permeable and liquid non-permeable membrane 22.
Figure 5. Illustrates results of a comparative study performed by an independent CRO for an emulsion prepared by the method of the present invention (·) and an emulsion prepared by the commonly used“2-syringe method” (O). The Experimental Autoimmune Encephalomyelitis (EAE) model, which is an animal disease model used to study the human disease Multiple Sclerosis, was used to compare the two different methods to prepare the emulsions for this study. Animals were injected with an emulsion consisting of Myelin protein (MOG) mixed with complete Freund’s Adjuvant (CFA) prepared using two different methods. Animals were scored daily for disease using the grades 0-8, depending on the degree of paralysis of each animal. Each group consisted of 10 animals. Shown are the mean disease scores ±SEM (standard error of mean). The induction of disease in the EAE model is dependent on the quality of the emulsion. Detailed description of the invention
The invention relates to a novel single device and method to prepare water-in oil and oil-in water emulsions, to induce immune responses in animals and humans. The combination of the device and the method will ensure rapid, consistent, and almost waste-free production of high quality emulsions. The invention is interrelated with: 1 ) induction of disease in autoimmune animal models, 2) testing potential immunotherapies of cancer treatments, 3) preparation of vaccines for use of immunization of humans and animals, and 4) immunization of animals for production of monoclonal and polyclonal antibodies for diagnostic and therapeutic purposes.
To induce satisfactory immune responses in the above examples stated, one needs an "adjuvant" (an immune enhancing substance) to be mixed with an antigen (a virus, bacteria, protein, peptides or haptens). The most commonly used oil adjuvant to create a water-in-oil emulsion is Freund’s adjuvant for immunizing animals. Using the present invention, a Perfect Oil & Water Emulsion Receptacle (POWER), together with the method described below, will generate an error-free, substance-saving, fast, consistent emulsion every time.
The main element of the invention is a single device 1 , that is to be used together with a shaking machine to prepare oil and water emulsions (water-in- oil and oil-in-water). The term emulsion may be interpreted as a dispersion of two liquids that are normally immiscible, whereas the term suspension may be interpreted as a heterogeneous mixture that contains solid particles. The device of the present invention may also be used to prepared suspensions. The device 1 , looks like a tube with one opening at one end 6, and a mating arrangement
7 at the other end. The mating arrangement 7 may be a snap-off closure 8. The mating arrangement may be fitted with other types of seals, such as cork, plug, stopper, screw cap, screw-lid, push-down lid, push-down plug, a peel-off seal, or with a plunger seal. To this device 1 , first adjuvants and antigens will be added by pipetting or other means, and then the first sealable opening 6, will be closed with a seal which may be a cork, plug, stopper, screw cap, screw-lid, push-down lid, push-down plug, a peel-off seal or with a plunger-seal. The device 1 , will then be shaken in a high speed "shaking machine" at a speed, for a time, depending on which machine is used to produce a perfect emulsion. In a perfect emulsion there is no distinguishable phase separation in the emulsion. To remove any air trapped in the emulsion, the device 1 , will be centrifuged for a short time at a low g-force. Finally, the emulsion will be transferred to a syringe.
In one embodiment the seal of the of the first sealable opening 6 of the device 1 will be removed and a plunger 12 with a plunger seal 13 attached will be used to push the emulsion into a syringe, or a plunger 14, with an outer thread 17 at one end, to be attached to a plunger seal 19 with an inner thread 18 with a short turn so that air can go through the threads 17 and 18, then it will be pushed in to the top of the emulsion to eliminate air trapped, then the plunger is turned so that it closes off the hole in the plunger seal 19. Further, a snap-off closure 8 is removed, and the back end of a syringe is fitted to the protrusion 9 and locked in with the hooks 16 on the device 1 , then the emulsion is pushed into the attached syringe. By plunger means that any plunger or plunger rod available and suitable for this purpose are included in the present invention.
In one embodiment of the invention, the device 1 , for preparing an emulsion, comprising a body 2 with a length 3, a width 4, and a cross section 5. Further, the body 2 comprise one first sealable opening 6, for introduction of components to be emulsified, and one opening comprising a snap-off closure 8. The protrusion 9 left after removal of the snap-off closure 8 together with the hooks 16 is designed to fit the back opening of a syringe. One purpose of the invention is to facilitate transfer of a created emulsion into a syringe without any trapped air.
In one embodiment the body 2 of the device 1 , is elongated. In another embodiment the body 2 is a tubular shape. In yet another embodiment the body 2 is a cylindrical shape, but also other geometrical shapes are thinkable. The device 1 , according to the present invention should not be seen as limited by a certain geometrical shape. Further, the device 1 , may be of various materials. In one embodiment the device 1 , is made of glass, metal and/or a polymer material. In one embodiment the polymer material is selected from thermoplastics or thermosetting polymers. In one example, to which the invention is not to be seen as limited, the present invention may be comprising, acrylic (polymethyl methacrylate, PMMA), Polycarbonate (PC), Polyvinyl chloride (PVC), Polypropylene (PP), Polyethylene terephthalate (PET / APET), PETG, Acrylonitrile Butadiene Styrene (ABS) and fluorinated polymers, such as fluorinated ethylene-propylene (FEP), polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF), perfluoroalkoxy alkane (PFA) and/or any combinations thereof.
When preparing an emulsion, it is desirable to contain the components to be emulsified. In one embodiment the device 1 comprising an opening 6, for introduction of components to be emulsified. The opening 6 will then be closed with a seal. Various seals are thinkable, examples of seals may be seals selected from the group consisting of screw lid, push lid, plug, thermoplastic plunger stopper, plunger seal, peel-off seal, cork or other devices that may seal one end of the device. In one embodiment, a plunger 12 with a plunger seal 13 or a plunger 14 with an outer thread 17, together with a plunger seal 19 with an inner thread 18, may be inserted into the first sealable opening 6 (the top) of the device 1 .
In one embodiment, the seal is a plunger head. In one embodiment the seal is adapted to mate with a plunger tip. In another embodiment the plunger comprises inner threads and the plunger tip outer threads. In yet another embodiment, the plunger 20 is fitted with a gas-permeable and liquid non- permeable membrane 22, which will be mounted into an opening at the end 21 . The end 21 has holes through the base where air can escape when the plunger is pushed into the syringe with emulsions.
In one embodiment the plunger-tip comprises a recess optionally comprising an opening with a narrower entrance portion to which a plunger with a protrusion at the end that can be fitted. A common seal is a screw lid. In one embodiment the body 2 of the device 1 of the present invention comprise threads 10. These threads 10 may be arranged in close proximity to the first sealable opening 6 of the device 1 where the components to be emulsified, are to be introduced. Further, the threads may be arranged to mate with the threads of a screw lid. Various angles of the threads are thinkable. In one embodiment the top of the device comprises an inner thread and the screw-lid comprise an outer thread. The outer thread of the screw lid is to be fitted with the inner thread of the device.
In one embodiment mating arrangement 7 of the device 1 , comprising hooks 16 and a protrusion 9, is adapted to fit the back end of a syringe.
In one embodiment the mating arrangement 7 fits an adaptor, for example a Luer Lock arrangement. This may be helpful for fitting syringes that are of larger sizes. Since one of the purposes of the present invention is to facilitate transfer of created emulsion, in one embodiment, the mating arrangement 7 comprise threads at the end, preferably outer threads.
In one embodiment the present invention comprises a female-to-female, male-to-male or male-to-female adaptor. However, this embodiment may result in some additional loss of products, compared to the embodiments not using this adaptor.
One application for emulsions may be, as mentioned, to be administered by a syringe. In one embodiment the mating arrangement 7 is adapted to mate with a syringe.
The device 1 according to the present invention may be of various sizes and is not to be seen as limited by a specific size. In one embodiment, the length 3 is larger than the width 4. In another embodiment the length 3 is selected from the group consisting of 10-500 mm, 10-300 mm, 10-200 mm, 10- 150 mm, 10-100 mm, 10-50 mm, and/or the width 4 is selected from the group consisting of 5-100 mm or 5-50 mm. In one embodiment, the relationship between the length 3 and the width 4 (lengtlrwidth) is at least 2:1 , at least 3:1 , at least 4:1 , at least 5:1 , at least 6:1 , at least 7:1 , at least 8:1 , at least 9:1 , at least 10:1 . In another embodiment the relationship between the length 3 and the width 4 is at most 15:1 . The device 1 may comprise various geometrical shapes. In one embodiment, the cross section 5, seen across the length 3 of the body 2 comprise a geometrical shape such as circular, oval, square or square with rounded corners, triangular, triangular with rounded corners. In another embodiment the cross section 5 comprises a geometrical shape with corners such as at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten corners. In one embodiment the cross section 5 comprises at most 20 corners.
When transferring the emulsion prepared from the device 1 , a force need to be applied to the plunger, in the direction along the body 2 of the device 1 and towards the end comprising the mating arrangement 7. To facilitate the application of this force, a flange 1 1 may be arranged in close proximity to the first sealable opening 6. The flange 1 1 may be a collar or a skirt. In one embodiment the flange 1 1 at least partially encloses the body 2 along part of the length 3. In another embodiment the flange 1 1 may be the shape of a ring, a collar, wings or similar to those on a syringe. Other shapes are thinkable and the purpose of the flange 1 1 is to stop the device from slipping through the finger while pushing the plunger into the device. The flange 1 1 is in one embodiment 1 -50 mm larger in radius than the cross section of the device 1 .
The device 1 may have different inner volumes. In one embodiment the device 1 comprises an inner volume, when sealed, of preferably 0.5-100 ml, preferably 1 -50 ml. In one embodiment the device 1 comprises an inner volume, selected from the group consisting of 0.5-5 ml, 1 -10 ml, 5-15 ml, 5-30 ml, 10-50 ml, 10-100 ml, and 50-500 ml.
In another aspect, the present invention is a method for creating an emulsion comprising the steps:
A8) filling the device 1 with the components intended to be emulsified;
B8) sealing the device 1 ;
C8) shaking the sealed device 1 to produce an emulsion.
In one embodiment the method further comprises a centrifugation step after shaking in step C8). In another embodiment the method further comprises removing the screw lid or push lid and inserting a plunger at the top, or inserting a plunger at the top containing an outer thread to fit the plunger-tip in the device with an inner thread, or inserting a plunger with a protrusion at the end to fit the plunger seal in the device comprising a recess opening with a narrower entrance portion, or inserting a plunger fitted with a gas-permeable, liquid non-permeable membrane at its end, and then removing the snap-off closure at the other end and attach a syringe.
In one embodiment step B8) of the present invention further comprises closing the first sealable opening 6 of the device 1 with a seal.
In another embodiment the method is further comprising the steps:
D8) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which comprises the sealable mating arrangement 7, wherein the mating arrangement 7 comprise a snap-off closure 8, further comprising hooks 16;
E8) removing the seal, of the first sealable opening 6 of the device 1 and inserting a syringe plunger, or inserting a plunger seal with an inner thread;
F8) removing snap-off closure 8, and attach a syringe and lock it in with the hooks 16 to fit the back opening of a syringe; and
G8) pushing the syringe plunger into the device 1 to force the emulsion through the second sealable opening 15 and into the attached syringe.
In another embodiment uses a plunger 14 containing a protrusion at the end with an outer thread 17 for pushing in a plunger seal 19 with an inner thread 18 to a close proximity of the surface of an emulsion without screwing in the plunger to let the air out, and then fastening the plunger to the plunger seal 19 before forcing the emulsion into an attached syringe.
In another embodiment the device 1 , looks like a tube with one opening at one end, a first sealable opening 6, and a female Luer Lock adapter at the other end, a second sealable opening 15. To this device 1 , first a male Luer Lock Cap will be attached to the female Luer Lock fitting, adjuvants and antigens will then be added by pipetting or other means, and finally the first sealable opening 6, is closed with a seal wherein the seal may be a screw cap, screw-lid, push-down lid, push-down plug or with a plunger seal and a screw cap. The device 1 , will then be shaken in a high speed "shaking machine" at a speed, for a time, depending on the machine used, to produce a perfect emulsion. The time for preparing an emulsion of high quality with the method of the present invention is preferably 0.5-12 min, preferably 1 -10 min, preferably, 2-8 min, preferably 5-7 min. In a perfect emulsion there is no distinguishable phase separation in the emulsion. To remove any air trapped in the emulsion, the device 1 , will be centrifuged for a short time at a low en force. Finally, the male Luer Lock Cap will be removed and any syringe of either male Luer Lock or Luer Slip type will be attached to the device 1 , screw lid removed and a plunger, attached to push the emulsion into the syringe, or a plunger, to be attached to the plunger seal 19 with a short turn so that air can go through the threads 18, then it is pushed in to a close proximity of the emulsion, then the plunger is turned so that it closes off the hole in plunger seal, and then the emulsion is pushed out into the attached syringe.
In one embodiment of the invention, the device 1 , for preparing an emulsion, comprising a body 2 with a length 3, a width 4, and a cross section 5. Further, the body 2 comprise a first sealable opening 6, for introduction of components to be emulsified, and a second sealable opening 15 comprising a female Luer Lock fitting. The female Luer lock fitting may be designed to fit a male Luer Lock fitting and/or a Slip fitting of a syringe. One purpose of the invention is to facilitate transfer of a created emulsion into a syringe, preferably under aseptic conditions.
In one embodiment the body 2 of the device 1 , is elongated. In another embodiment the body 2 is a tubular shape. In yet another embodiment the body 2 is a cylindrical shape, but also other geometrical shapes are thinkable. The device 1 , according to the present invention should not be seen as limited by a certain geometrical shape.
When preparing an emulsion, it is desirable to contain the components to be emulsified. In one embodiment the device 1 , further comprises a seal, arranged in close proximity to the first sealable opening 6, for introduction of components to be emulsified. In one embodiment the first sealable opening 6 is a sealable end. The sealable end 6 may be sealed with a seal. Various seals are thinkable, examples of seals may be seals selected from the group consisting of screw lid, push lid, plug, stopper, plunger-tip, cork or other devices that may seal one end of the device. In the present application the wording seal and close are used interchangeably.
In one embodiment, a plunger seal 13 and/or a plunger seal 19, with an inner thread 18, through the hole of plunger seal, may be inserted into the first opening 6 of the device 1 . In this embodiment, the seal may further comprise a plunger 14, with outer threads 17 at the end that can be fitted with the plunger seal 19 with inner threads 18. In one embodiment, the seal is a plunger head.
In one embodiment the plunger seal comprises a recess optionally comprising an opening with a narrower entrance portion to which a plunger with a protrusion at the end that can be fitted.
A common seal for the first sealable opening 6 is a screw lid. In one embodiment the body 2 of the device 1 of the present invention comprise threads 10. These threads may be arranged in close proximity to the first sealable opening 6 of the device 1 where the components to be emulsified, are to be introduced. Further, the threads may be arranged to mate with the threads of a screw lid. Various angles of the threads are thinkable. In one embodiment the top of the device comprises an inner thread and the screw-lid comprise an outer thread. The outer thread of the screw lid is to be fitted with the inner thread of the device.
In one embodiment mating arrangement 7 of the device is adapted to fit male Luer lock fittings preferably of syringes.
In one embodiment the mating arrangement 7 fits an adaptor. This may be helpful for fitting syringes that lack Luer lock. Since one of the purposes of the present invention is to facilitate transfer of created emulsion, in one embodiment, the mating arrangement 7 comprise threads at the end, preferably outer threads.
In one embodiment the present invention comprises a female-to-female, male-to-male or male-to-female adaptor. However, this embodiment may result in some additional loss of products compared to not using this adaptor. One application for emulsions may be, as mentioned, to be administered by a syringe. In one embodiment the mating arrangement 7 is adapted to mate with a syringe.
The device 1 according to the present invention may be of various sizes and is not to be seen as limited by a specific size. In one embodiment, the length 3 is larger than the width 4. In another embodiment the length 3 is selected from the group consisting of 10-500 mm, 10-300 mm, 10-200 mm, 10- 150 mm, 10-100 mm, 10-50 mm, and/or the width 4 is selected from the group consisting of 5-100 mm or 5-50 mm. In one embodiment, the relationship between the length 3 and the width 4 (lengtlrwidth) is at least 2:1 , at least 3:1 , at least 4:1 , at least 5:1 , at least 6:1 , at least 7:1 , at least 8:1 , at least 9:1 , at least 10:1 . In another embodiment the relationship between the length 3 and the width 4 is at most 15:1 .
The device 1 may comprise various geometrical shapes. In one embodiment, the cross section 5, seen across the length 3 of the body 2 comprise a geometrical shape such as circular, oval, square or square with rounded corners, triangular, triangular with rounded corners. In another embodiment the cross section 5 comprises a geometrical shape with corners such as at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten corners. In one embodiment the cross section 5 comprises at most 20 corners.
When transferring the emulsion prepared from the device 1 , a force need to be applied to the plunger 12, in the direction along the body 2 of the device 1 and towards the second sealable opening 15 comprising the mating arrangement 7. To facilitate the application of this force, a flange 1 1 may be arranged in close proximity to the first sealable opening 6. The flange 1 1 may be a collar or a skirt. In one embodiment the flange 1 1 at least partially encloses the body 2 along part of the length 3. In another embodiment the collar may be the shape of a ring, a flange, wings or similar to those on a syringe. Other shapes are thinkable and the purpose of the flange 1 1 is to stop the device from slipping through the finger while pushing the plunger into the device. The flange 1 1 is in one embodiment 1 -50 mm larger in radius than the cross section of the device 1 .
The device 1 may have different inner volumes. In one embodiment the device 1 comprises an inner volume, when sealed, of preferably 0.5-100 ml, preferably 1 -50 ml. In one embodiment the device 1 comprises an inner volume, selected from the group consisting of 0.5-5 ml, 1 -10 ml, 5-15 ml, 5-30 ml, 10-50 ml, 10-100 ml, and 50-500 ml.
In another aspect, the present invention is a method for creating an emulsion comprising the steps:
A9) attaching a stop plug to the mating arrangement 7 near the second sealable opening 15 of the device 1 according to the present invention;
B9) filling the device 1 with the components intended to be emulsified;
C9) closing the device 1 with a seal;
D9) shaking the sealed device 1 to produce an emulsion.
In one embodiment the method further comprises a centrifugation step after shaking in step D9).
In one embodiment the mating arrangement 7 may be a female Luer Lock fitting.
In another embodiment the method further comprises removing the screw lid or push lid and inserting a plunger at the top, or inserting a plunger at the top containing an outer thread to fit the plunger seal in the device with an inner thread, or inserting a plunger with a protrusion at the end to fit the plunger seal in the device comprising a recess opening with a narrower entrance portion, or inserting a plunger fitted with a gas-permeable, liquid non-permeable membrane at its end, and then removing the stop plug at the other end and attach a male Luer lock or Luer slip syringe.
Another aspect of the present invention relates to a method for creating an emulsion comprising the steps:
A10) filling the device 1 with the components intended to be emulsified;
B10) closing the first sealable opening 6 of the device 1 with a seal; and
C10) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps: D10) centrifugation and/or hand shaking of the device 1 to force the emulsion towards a sealed second sealable opening 15 of the device 1 , which second sealable opening 15 comprise the mating arrangement 7;
E10) removing the seal, of the first sealable opening 6 of the device 1 and inserting a syringe plunger;
F10) connecting the back end opening of a syringe, without its plunger, to the mating arrangement 7; and
G10) pushing the syringe plunger into the device 1 to force the emulsion through the second sealable opening 15 and into the connected syringe. Another aspect of the present invention relates to a method for creating an emulsion comprising the steps:
A11) attaching a stop plug to the female Luer Lock fitting at the second sealable opening of the device 1 according to the present invention;
B11) filling the device 1 with the components intended to be emulsified;
C11) closing the first sealable opening 6 of the device 1 with a seal;
D11) shaking the sealed device 1 to produce an emulsion.
In another embodiment the method is further comprising the steps:
E11) centrifugation and/or hand shaking of the device 1 to force the emulsion towards the second sealable opening 15 of the device 1 , which second sealable opening comprises the mating arrangement 7;
F11) connecting a male Luer Lock or Luer Slip syringe to the mating arrangement 7;
G11) removing the seal, of the first sealable opening 6 of the device 1 and inserting a syringe plunger, or inserting a plunger-tip with an inner thread; and
H11) pushing the syringe plunger into the device 1 to force the emulsion through the second sealable opening 15 and into a connected syringe. In one embodiment, the method further comprises a step to use a plunger containing a protrusion at the end with an outer thread to push in the plunger seal with an inner thread to close proximity of the surface of the emulsion without screwing in the plunger to let the air out, and then fasten the plunger to the plunger seal before forcing the emulsion through the second sealable opening and into an attached syringe.
In yet another embodiment, the method further comprise pushing out the emulsion into the syringe by pressing in the plunger at the top and stopping the device to move by holding onto the protrusion. Another aspect of the present invention relates to a kit of parts comprising the device according to the present invention, and an instruction pamphlet comprising the method according to what is disclosed herein, regarding the present invention.
In yet another aspect, the present invention relates to use of the device according to what is disclosed herein for production of an emulsion.
And in yet another aspect, the invention relates to an emulsion obtained according to the method of the present invention, described herein.
In one embodiment the present invention is suitable for preparing liposomes.
Examples
The device and method described here have a number of advantages.
1 ) no person to person variation making the emulsion
2) almost no waste of expensive material. With the various syringe methods (for example 2-way and 3-way), up to 50% of the emulsion is lost during preparation, and
3) it is fast. Making an emulsion of high quality takes approximately 6 minutes, compare to approximately 15 minutes for the syringe method. An example of comparing the method in the present application with the common 2-syringe method. Interesting result is that upon performing an animal experiment where the quality of the emulsion is very important.
Figure 5 shows results of a comparative study for an emulsion prepared by the method of the present invention and a control emulsion, by using the Experimental Autoimmune Encephalomyelitis (EAE) model, which is an animal disease model used to study the human disease Multiple Sclerosis. Preparation of a high-quality water-in-oil emulsion containing Myelin protein is necessary to induce disease in research animals. Animals were injected with an emulsion consisting of Myelin protein (MOG) mixed with complete Freund’s Adjuvant (CFA) prepared using two different methods. A standard protocol using the 2- syringe method (O) with an emulsion that was prepared using the method described by the present invention (·). Animals were scored daily for disease using the grades 0-8, depending on the degree of paralysis of each animal. Each group consisted of 10 animals. Shown are the mean disease scores ±SEM (standard error of mean).
As is evident from the above, the present invention provides a homogenous emulsion that is independent on personal differences. The common 2-syringe method varies from person to person and can be of very different quality, depending on finger strength, time and speed used, temperature in the laboratory, just mentioning a few parameters. When prepared using the 2-syringe method a large proportion of the emulsion is lost in the parts used. In the present invention, there is almost no loss of emulsion except the dead volume in the syringe used. Flowever, this loss is not specific to the present invention. The same loss will also occur when using the 2-syringe method. It should be noted that the device according to the present invention of course may comprise different standard components, also having varied design, and being manufactured in various thinkable materials which are not explicitly mentioned. Moreover, the design of the device may vary, and the present invention, as formulated in the present claim set, should be seen as embodying different forms and materials of the device.

Claims

Claims
1. Device (1 ) adapted for preparing an emulsion therein, comprising a body (2) with a length (3), a width (4), and a cross section (5), further comprising a first sealable opening (6), for introduction of components to be emulsified, a second sealable opening (15) comprising a mating arrangement (7), wherein the mating arrangement (7) is adapted to mate with an external part.
2. Device (1 ) according to claim 1 , wherein the mating arrangement (7) is adapted to mate with a syringe.
3. Device (1 ) according to claim 1 or 2, wherein the mating arrangement (7) and the first sealable opening (6) are arranged on opposite sides of the body (2).
4. Device (1 ) according to any one of claims 1 -3, wherein the body (2) is elongated, preferably in a tubular shape, preferably made of a material selected from the group consisting of glass, metal and a polymer material, preferably selected from thermoplastics or thermosetting polymers.
5. Device (1 ) according to any one of claims 1 -4, further comprising a flange (1 1 ), preferably arranged in close proximity to the first sealable opening (6).
6. Device (1 ) according to any one of claims 1 -5, comprising an inner volume, when sealed, of 0.5-500 ml, preferably 1 -500 ml, preferably 1 -100 ml, preferably 1 -50 ml, preferably 1 -10 ml, preferably 5-10 ml.
7. Device (1 ) according to any one of claims 1 -6, wherein the mating arrangement (7) comprises a protrusion (9), preferably the protrusion (9) comprises a snap-off closure (8). 8. Device (1 ) according to any one of claims 1 -7, comprising a plunger
(14) adapted for being inserted through the first sealable opening (6), wherein the plunger comprises a protrusion at the end with an outer thread (17) to fit a plunger seal (19) with an inner thread (18). 9. Device (1 ) according to any one of claims 1 -7, comprising a plunger
(20), with an opening at the end (21 ), to fit a gas-permeable, liquid non-permeable membrane (22).
10. Method for creating an emulsion comprising the steps:
A) filling the device (1 ) according to claim 1 with the components intended to be emulsified;
B) closing the first sealable opening (6) of the device (1 ); and
C) shaking the closed device (1 ) to produce an emulsion. 1 1 . Method according to claim (10), further comprising the steps:
D) centrifugation and/or hand shaking of the device (1 ) to force the emulsion towards the second sealable opening (15) comprising the mating arrangement (7);
E) connecting a syringe to the mating arrangement (7);
F) forcing the emulsion from the device (1 ), through the mating arrangement (7), and into the connected syringe.
12. Use of the device (1 ) according to claim 1 -9 for production of an emulsion.
13. Emulsion obtained according to the method of any one of claims 10- 1 1 .
PCT/SE2019/050752 2018-08-17 2019-08-16 Device and method for preparing water-in-oil and oil-in-water emulsions for immunizations of animals and humans WO2020036531A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP19849419.7A EP3836884A4 (en) 2018-08-17 2019-08-16 Device and method for preparing water-in-oil and oil-in-water emulsions for immunizations of animals and humans

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE1850986 2018-08-17
SE1850986-9 2018-08-17

Publications (1)

Publication Number Publication Date
WO2020036531A1 true WO2020036531A1 (en) 2020-02-20

Family

ID=69525755

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE2019/050752 WO2020036531A1 (en) 2018-08-17 2019-08-16 Device and method for preparing water-in-oil and oil-in-water emulsions for immunizations of animals and humans

Country Status (2)

Country Link
EP (1) EP3836884A4 (en)
WO (1) WO2020036531A1 (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1466572A2 (en) * 2003-03-04 2004-10-13 Sidam di Azzolini Graziano E C. S.a.s. Device for packaging, mixing and applying bone cement
WO2007068032A1 (en) * 2005-12-12 2007-06-21 Nature Vet Pty Ltd An apparatus for holding and dispensing flowable preparations
US20080044606A1 (en) * 2004-09-27 2008-02-21 Kenji Omori Medical Container and Medical Duplex Container
US20110166596A1 (en) * 2010-01-05 2011-07-07 Baxter International Inc. Mixing System, Kit and Mixer Adapter
US20120089088A1 (en) * 2009-06-02 2012-04-12 Foshee David L Multi-container transfer and delivery device
US20150265500A1 (en) * 2012-12-17 2015-09-24 Robert Scott Russo Vial adapters

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE20008103U1 (en) * 2000-05-05 2001-09-13 Coripharm Medizinprodukte Gmbh Preparation and application device for materials, in particular bone cement, to be prepared from at least one powdery or granular and a liquid component immediately before use to form a pasty, flowable mass

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1466572A2 (en) * 2003-03-04 2004-10-13 Sidam di Azzolini Graziano E C. S.a.s. Device for packaging, mixing and applying bone cement
US20080044606A1 (en) * 2004-09-27 2008-02-21 Kenji Omori Medical Container and Medical Duplex Container
WO2007068032A1 (en) * 2005-12-12 2007-06-21 Nature Vet Pty Ltd An apparatus for holding and dispensing flowable preparations
US20120089088A1 (en) * 2009-06-02 2012-04-12 Foshee David L Multi-container transfer and delivery device
US20110166596A1 (en) * 2010-01-05 2011-07-07 Baxter International Inc. Mixing System, Kit and Mixer Adapter
US20150265500A1 (en) * 2012-12-17 2015-09-24 Robert Scott Russo Vial adapters

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP3836884A4 *

Also Published As

Publication number Publication date
EP3836884A4 (en) 2022-05-11
EP3836884A1 (en) 2021-06-23

Similar Documents

Publication Publication Date Title
JP5385383B2 (en) Density phase separator
US9399226B2 (en) Method and apparatus for separating fluid components
US20240058807A1 (en) Isolation tube
US7914689B2 (en) Apparatus and method for separating and concentrating fluids containing multiple components
US20220072489A1 (en) Mixing Element for Container Assemblies
US10987672B2 (en) Dual piston centrifuge tube
AU2009221836B2 (en) Co-molded pierceable stopper and method for making the same
JPS63500679A (en) Method and apparatus for separating serum/plasma from blood
EP0311011A2 (en) Device for separating the components of a liquid sample having higher and lower specific gravities
EP3142764B1 (en) System and apparatus for isolating or enriching agents using floatation
EP3847965A1 (en) Specimen container and method for separating serum or plasma from whole blood
JP2001264316A (en) Device and method for collecting, preparing and stabilizing sample
JP2000189407A (en) Assembly for component separation of fluid sample
RU2692064C2 (en) Container
EP3383269B1 (en) Apparatus for preparing blood fraction concentrate
JP2024010236A (en) Septum that decontaminates by interaction with penetrating element
WO2020036531A1 (en) Device and method for preparing water-in-oil and oil-in-water emulsions for immunizations of animals and humans
JPH01284348A (en) Releasable lock, assembly with the lock and method for its use
TWM614848U (en) Blood separation device
JPS584593Y2 (en) centrifuge tube
US20220244237A1 (en) System and device for separating or mixing biological fluids and/or tissue
KR101333788B1 (en) Separating vials and their uses
JPS61293467A (en) Gasket body for medical instrument
TW202241351A (en) Blood separation device

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19849419

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2019849419

Country of ref document: EP

Effective date: 20210317