WO2024153916A2

WO2024153916A2 - Systems and methods for aseptic connection

Info

Publication number: WO2024153916A2
Application number: PCT/GB2024/050092
Authority: WO
Inventors: James Davies; Antoine ESPINET
Original assignee: Microfluidx Ltd
Priority date: 2023-01-16
Filing date: 2024-01-15
Publication date: 2024-07-25
Also published as: WO2024153916A3

Abstract

The present disclosure provides systems, devices, and methods for aseptic connection. In some cases, a sterilization apparatus described herein comprises: (a) a sterilization chamber, (b) a first connector valve comprising a first fluid line embedded therein, wherein said first connector valve forms a fluid tight seal against said sterilization chamber; (c) a second connector valve comprising a second fluid line embedded therein, wherein said second connector valve forms a fluid tight seal against said sterilization chamber, wherein said second connector valve is configured to move from a first position to a second position.

Description

SYSTEMS AND METHODS FOR ASEPTIC CONNECTION

CROSS REFERENCE

[0001] This application claims priority to U.S. Provisional Patent Application No. 63/439,268, filed on January 16, 2023, which is incorporated herein by reference in entirety.

BACKGROUND

[0002] Ex-vivo processing of cells or other biological components for the research and development of cell therapy treatments requires a sterile environment to prevent the cells becoming infected by microbial growth. Given the complexity of processing, multiple fluid reagents must be delivered to multiple separate bioreactors throughout an experiment. Reagent tanks and bioreactors therefore must be able to fluidically connect and disconnect from each other, all without breaching sterility. [0003] Standard sterile aseptic connectors used to fluidly connect two sterile systems are typically disposable. In general, these connectors are normally single use and are typically designed in such a way that the two halves of the connectors come together at a pair of shutters which have a sterile and a dirty side. Upon joining the two halves, the two dirty sides of the shutters come into contact with one another. The shutters are then designed to be pulled away from the joined connectors, and in the process, sterile seals behind the shutters come into contact with each other and maintain the sterile barrier.

[0004] A common problem with these connectors is that by design they are single use and their unit cost is typically relatively high. Using them in a system with a high number of bioreactors and or reagent vessels results in the use of many aseptic connectors which results in a high consumable cost of the machine and lots of waste.

[0005] Thus, there is a need for reusable methods and systems for fluidically connecting two or more sterile systems without breaching sterility. SUMMARY

[0006] The systems and methods described herein disclose a reusable alternative to standard aseptic that can be used to connect two or more separate sterile systems (i.e., reagent tanks, bioreactors, cassettes, valves, pumps) such that that one continuous sterile barrier is formed that encompasses all systems.

[0007] In an aspect, provided herein is a sterilization apparatus, comprising: (a) a sterilization chamber, (b) a first connector valve comprising a first fluid line embedded therein, wherein the first connector valve forms a fluid tight seal against the sterilization chamber; and (c) a second connector valve comprising a second fluid line embedded therein, wherein the second connector valve forms a fluid tight seal against the sterilization chamber, wherein the second connector valve is configured to move from a first position to a second position, wherein when the second connector valve is in the first position, the first connector valve and the second connector valve are closed such that the sterilization chamber is fluidically sealed from the first fluid line and the second fluid line, and wherein when the second connector valve is in the second position, the second connector valve engages with the first connector valve, thereby opening the first connector valve and the second connector valve such that the first fluid line and the second fluid line are fluidically connected.

[0008] In some cases, the first connector valve comprises a first hollow connector body with a first diameter, and the first hollow connector body is fluidically connected to the first fluid line. In some cases, the first connector valve comprises a first connector pin with a second diameter, and the second diameter is greater than the first diameter. In some cases, the first connector pin is configured to move from an extended position to a retracted position, and when in the extended position, at least a portion of the first connector pin extends into the sterilization chamber, and when in the retracted position, the first connector pin retracts into the first hollow connector body. In some cases, the at least portion of the first connector pin has a first fluid channel embedded therein. In some cases, when the first connector pin is in the extended position, the first fluid channel is located entirely within the sterilization chamber. In some cases, when the first connector pin is in the extended position, the first fluid channel is fluidically sealed from the first fluid line, thereby resulting in the first connector valve being closed.

[0009] In some cases, the first connector pin comprises a first front connector pin seal, and when the first connector pin is in the extended position, the first front connector pin seal causes the first fluid channel to be fluidically sealed from the first fluid line, thereby resulting in the first connector valve being closed. In some cases, when the first connector pin is in the retracted position, the first fluid channel is fluidically connected to the first fluid line, thereby resulting in the first connector valve being open. In some cases, when the second connector valve is in the first position, the first connector pin is in the extended position. In some cases, when the second connector valve is in the second position, the first connector pin is in the retracted position.

[0010] In some cases, the first connector valve further comprises a first compression spring, the first compression spring being configured to move from a first state to a second state. In some cases, the first compression spring is compressed more in the second state than the first state, and when the second connector valve is in the first position, the first compression spring is in the first state, and when the second connector valve is in the second position, the first compression spring is in the second state. In some cases, the first connector pin comprises a first back connector pin seal configured to fluidically seal the first fluid line from the first compression spring.

[0011] In some cases, the second connector valve comprises a second hollow connector body with a first diameter. In some cases, the second hollow connector body is fluidically connected to the second fluid line. In some cases, the second connector valve comprises a second connector pin with a second diameter, and the first diameter is greater than the second diameter. In some cases, the second connector pin is configured to move from an extended position to a retracted position, In some cases, when in the extended position, at least a portion of the second connector pin extends into the sterilization chamber, and when in the retracted position, the second connector pin retracts into the second hollow connector body. In some cases, the at least portion of the second connector pin has a second fluid channel embedded therein. In some cases, when the second connector pin is in the extended position, the second fluid channel is located entirely within the sterilization chamber. In some cases, when the second connector pin is in the extended position, the second fluid channel is fluidically sealed from the second fluid line, thereby resulting in the second connector valve being closed. In some cases, the second connector pin comprises a second front connector pin seal, and when the second connector pin is in the extended position, the second front connector pin seal causes the second fluid channel to be fluidically sealed from the second fluid line, thereby resulting in the second connector valve being closed. In some cases, when the second connector pin is in the retracted position, the second fluid channel is fluidically connected to the second fluid line, thereby resulting in the second connector valve being open.

[0012] In some cases, when the second connector valve is in the first position, the second connector pin is in the extended position. In some cases, when the second connector valve is in the second position, the second connector pin is in the retracted position.

[0013] In some cases, the second connector valve further comprises a second compression spring, and the second compression spring is configured to move from a first state to a second state. In some cases, the second compression spring is compressed more in the second state than the first state, and when the second connector valve is in the first position, the second compression spring is in the first state, and when the second connector valve is in the second position, the second compression spring is in the second state. In some cases, the second connector pin comprises a second back connector pin seal configured to fluidically seal the second fluid line from the second compression spring.

[0014] In some cases, the first fluid line is configured to fluidically connect to one or more reagent containers, and the second fluid line is configured to fluidically connect to one or more bioprocessing chambers. In some cases, the second fluid line is configured to fluidically connect to one or more reagent containers, and the first fluid line is configured to fluidically connect to one or more bioprocessing chambers. In some cases, the sterilization chamber is configured to receive a sterilization fluid. In some cases, the sterilization fluid comprises steam. In some cases, the sterilization fluid is configured to sterilize interior surfaces of the sterilization chamber. In some cases, a sterilization fluid is configured to sterilize the sterilization chamber and the first fluid channel when the first connector pin is in the extended position.

[0015] In some cases, the sterilization box comprises one or more of the first connector valves and one or more of the second connector valves, and each of the one or more first connector valves are paired with each of the one or more second connector valves.

[0016] In some cases, the sterilization box further comprises a latch, and the latch is configured to move the second connector valve from the first position to the second position. In some cases, the latch is configured to be moved manually by one or more users. In some cases, the latch is configured to be moved automatically by one or more mechanical elements.

[0017] In some cases, when the second connector valve is in the second position, the second connector valve at least partially surrounds a portion of the first connector valve. In some cases, the first connector valve comprises a male connector. In some cases, the second connector valve comprises a female connector. In some cases, when the second connector valve is in the second position, the first fluid line and the second fluid line are fluidically sealed from the sterilization chamber.

[0018] In another aspect, provided herein is a method, comprising: (a) providing a sterilization box comprising: (i) a sterilization chamber, (ii) a first connector valve comprising a first fluid line embedded therein, and (iii) a second connector valve comprising a second fluid line embedded therein, wherein the second connector valve is configured to move from a first position to a second position, (b) while the second connector valve is in the first position, introducing a sterilization fluid into the sterilization chamber, wherein when the connector valve is in the first position, the first connector valve and the second connector valve are closed such that the sterilization chamber is fluidically sealed from the first fluid channel and the second fluid channel, and the first connector valve and the second connector valve form a fluid tight seal against the sterilization chamber; and (c) adjusting a position of the second connector valve from the first position to the second position such that the second connector valve engages with the first connector valve, thereby resulting in the first fluid channel and the second fluid channel being fluidically connected, wherein the first fluid channel and the second fluid channel form a sterile fluid pathway.

[0019] In some cases, the first connector valve comprises a first hollow connector body with a first diameter, and the first hollow connector body is fluidically connected to the first fluid line. In some cases, the first connector valve comprises a first connector pin with a second diameter, and the second diameter is greater than the first diameter.

[0020] In some cases, the first connector pin is configured to move from an extended position to a retracted position, and when in the extended position, at least a portion of the first connector pin extends into the sterilization chamber. In some cases, when in the retracted position, the first connector pin retracts into the first hollow connector body.

[0021] In some cases, the at least portion of the first connector pin has a first fluid channel embedded therein. In some cases, when the first connector pin is in the extended position, the first fluid channel is located entirely within the sterilization chamber. In some cases, when the first connector pin is in the extended position, the first fluid channel is fluidically sealed from the first fluid line, thereby resulting in the first connector valve being closed.

[0022] In some cases, the first connector pin comprises a first front connector pin seal, and when the first connector pin is in the extended position, the first front connector pin seal causes the first fluid channel to be fluidically sealed from the first fluid line, thereby resulting in the first connector valve being closed.

[0023] In some cases, when the first connector pin is in the retracted position, the first fluid channel is fluidically connected to the first fluid line, thereby resulting in the first connector valve being open. In some cases, when the second connector valve is in the first position, the first connector pin is in the extended position. In some cases, when the second connector valve is in the second position, the first connector pin is in the retracted position.

[0024] In some cases, the first connector valve further comprises a first compression spring, the first compression spring being configured to move from a first state to a second state. In some cases, the first compression spring is compressed more in the second state than the first state, and when the second connector valve is in the first position, the first compression spring is in the first state, and when the second connector valve is in the second position, the first compression spring is in the second state. In some cases, the first connector pin comprises a first back connector pin seal configured to fluidically seal the first fluid line from the first compression spring.

[0025] In some cases, the second connector valve comprises a second hollow connector body with a first diameter, and the second hollow connector body is fluidically connected to the second fluid line. In some cases, the second connector valve comprises a second connector pin with a second diameter, and the first diameter is greater than the second diameter. In some cases, the second connector pin is configured to move from an extended position to a retracted position, and when in the extended position, at least a portion of the second connector pin extends into the sterilization chamber, and when in the retracted position, the second connector pin retracts into the second hollow connector body.

[0026] In some cases, the at least portion of the second connector pin has a second fluid channel embedded therein. In some cases, when the second connector pin is in the extended position, the second fluid channel is located entirely within the sterilization chamber. In some cases, when the second connector pin is in the extended position, the second fluid channel is fluidically sealed from the second fluid line, thereby resulting in the second connector valve being closed.

[0027] In some cases, the second connector pin comprises a second front connector pin seal, and when the second connector pin is in the extended position, the second front connector pin seal causes the second fluid channel to be fluidically sealed from the second fluid line, thereby resulting in the second connector valve being closed.

[0028] In some cases, when the second connector pin is in the retracted position, the second fluid channel is fluidically connected to the second fluid line, thereby resulting in the second connector valve being open. In some cases, when the second connector valve is in the first position, the second connector pin is in the extended position. In some cases, when the second connector valve is in the second position, the second connector pin is in the retracted position.

[0029] In some cases, the second connector valve further comprises a second compression spring, the second compression spring being configured to move from a first state to a second state. In some cases, the second compression spring is compressed more in the second state than the first state. In some cases, when the second connector valve is in the first position, the second compression spring is in the first state, and when the second connector valve is in the second position, the second compression spring is in the second state. In some cases, the second connector pin comprises a second back connector pin seal configured to fluidically seal the second fluid line from the compression spring.

[0030] In some cases, the first fluid line is configured to fluidically connect to one or more reagent containers, and the second fluid line is configured to fluidically connect to one or more bioprocessing chambers. In some cases, the second fluid line is configured to fluidically connect to one or more reagent containers, and the first fluid line is configured to fluidically connect to one or more bioprocessing chambers.

[0031] In some cases, the sterilization fluid comprises steam. In some cases, the sterilization fluid is configured to sterilize interior surfaces of the sterilization chamber. In some cases, the sterilization fluid is configured to sterilize the sterilization chamber and the first fluid channel when the first connector pin is in the extended position. In some cases, the sterilization fluid is configured to sterilize the sterilization chamber and the second fluid channel when the second connector pin is in the extended position.

[0032] In some cases, the sterilization box comprises one or more of the first connector valves and one or more of the second connector valves, and each of the one or more first connector valves are paired with each of the one or more second connector valves.

[0033] In some cases, the sterilization box further comprises a latch, and the latch is configured to perform the adjusting in (c). In some cases, the latch is configured to be adjusted manually by one or more users. In some cases, the latch is configured to be adjusted automatically by one or more mechanical elements.

[0034] In some cases, when the second connector valve is in the second position, the second connector valve at least partially surrounds a portion of the first connector valve. In some cases, the first connector valve comprises a male connector. In some cases, the second connector valve comprises a female connector. In some cases, when the second connector valve is in the second position, the first fluid line and the second fluid line are fluidically sealed from the sterilization chamber.

[0035] In another aspect, provided herein is a microfluidic system, comprising: (a) one or more reagent containers; (b) one or more bioprocessing chambers; and (c) a sterilization apparatus comprising (i) a sterilization chamber, (ii) a first connector valve fluidically connected to the one or more reagent containers, wherein the first connector valve forms a fluid tight seal against the sterilization chamber, and (iii) a second connector fluidically connected to the one or more bioprocessing chambers, wherein the second connector valve forms a fluid tight seal against the sterilization chamber; wherein the first connector valve or the second connector valve is configured to move from a first position to a second position, wherein when the first connector valve or the second connector valve is in the first position, the first connector valve and the second connector valve are closed such that the sterilization chamber is fluidically sealed from the one or more reagent containers and the one or more bioprocessing chambers, and wherein when the first connector valve or the SECOND connector valve is in the second position, the second connector valve engages with the first connector valve, thereby opening the first connector valve and the second connector valve such that the one or more reagent containers and the one or more bioprocessing chambers are fluidically connected.

[0036] In some cases, the first connector valve comprises a first hollow connector body with a first diameter, and the first hollow connector body is fluidically connected to the first fluid line. In some cases, the first connector valve comprises a first connector pin with a second diameter, and the second diameter is greater than the first diameter. In some cases, the first connector pin is configured to move from an extended position to a retracted position, and when in the extended position, at least a portion of the first connector pin extends into the sterilization chamber, and when in the retracted position, the first connector pin retracts into the first hollow connector body.

[0037] In some cases, the at least portion of the first connector pin has a first fluid channel embedded therein. In some cases, when the first connector pin is in the extended position, the first fluid channel is located entirely within the sterilization chamber. In some cases, when the first connector pin is in the extended position, the first fluid channel is fluidically sealed from the first fluid line, thereby resulting in the first connector valve being closed. In some cases, the first connector pin comprises a first front connector pin seal, and when the first connector pin is in the extended position, the first front connector pin seal causes the first fluid channel to be fluidically sealed from the first fluid line, thereby resulting in the first connector valve being closed.

[0038] In some cases, when the first connector pin is in the retracted position, the first fluid channel is fluidically connected to the first fluid line, thereby resulting in the first connector valve being open. In some cases, when the second connector valve is in the first position, the first connector pin is in the extended position. In some cases, when the second connector valve is in the second position, the first connector pin is in the retracted position.

[0039] In some cases, the first connector valve further comprises a first compression spring, and the first compression spring is configured to move from a first state to a second state. In some cases, the first compression spring is compressed more in the second state than the first state. In some cases, when the second connector valve is in the first position, the first compression spring is in the first state, and when the second connector valve is in the second position, the first compression spring is in the second state. In some cases, the first connector pin comprises a first back connector pin seal configured to fluidically seal the first fluid line from the first compression spring.

[0040] In some cases, the second connector valve comprises a second hollow connector body with a first diameter, the second hollow connector body being fluidically connected to the second fluid line. In some cases, the second connector valve comprises a second connector pin with a second diameter, and the first diameter is greater than the second diameter.

[0041] In some cases, the second connector pin is configured to move from an extended position to a retracted position. In some cases, when in the extended position, at least a portion of the second connector pin extends into the sterilization chamber, and when in the retracted position, the second connector pin retracts into the second hollow connector body.

[0042] In some cases, the at least portion of the second connector pin has a second fluid channel embedded therein. In some cases, when the second connector pin is in the extended position, the second fluid channel is located entirely within the sterilization chamber. In some cases, when the second connector pin is in the extended position, the second fluid channel is fluidically sealed from the second fluid line, thereby resulting in the second connector valve being closed.

[0043] In some cases, the second connector pin comprises a second front connector pin seal, and when the second connector pin is in the extended position, the second front connector pin seal causes the second fluid channel to be fluidically sealed from the second fluid line, thereby resulting in the second connector valve being closed. In some cases, when the second connector pin is in the retracted position, the second fluid channel is fluidically connected to the second fluid line, thereby resulting in the second connector valve being open. In some cases, when the second connector valve is in the first position, the second connector pin is in the extended position. In some cases, when the second connector valve is in the second position, the second connector pin is in the retracted position.

[0044] In some cases, the second connector valve further comprises a second compression spring, the second compression spring being configured to move from a first state to a second state. In some cases, the second compression spring is compressed more in the second state than the first state. In some cases, when the second connector valve is in the first position, the second compression spring is in the first state, and when the second connector valve is in the second position, the second compression spring is in the second state.

[0045] In some cases, the second connector pin comprises a second back connector pin seal configured to fluidically seal the second fluid line from the compression spring.

[0046] In some cases, the sterilization chamber is configured to receive a sterilization fluid. In some cases, the sterilization fluid comprises steam. In some cases, the sterilization fluid is configured to sterilize interior surfaces of the sterilization chamber. In some cases, the sterilization fluid is configured to sterilize the sterilization chamber and the first fluid channel when the first connector pin is in the extended position. In some cases, the sterilization fluid is configured to sterilize the sterilization chamber and the second fluid channel when the second connector pin is in the extended position.

[0047] In some cases, the sterilization box comprises one or more of the first connector valves and one or more of the second connector valves, and each of the one or more first connector valves are paired with each of the one or more second connector valves.

[0048] In some cases, the sterilization box further comprises a latch, and the latch is configured to move the second connector valve from the first position to the second position. In some cases, the latch is configured to be adjusted manually by one or more users. In some cases, the latch is configured to be adjusted automatically by one or more mechanical elements.

[0049] In some cases, when the second connector valve is in the second position, the second connector valve at least partially surrounds a portion of the first connector valve. In some cases, the first connector valve comprises a male connector. In some cases, the second connector valve comprises a female connector.

[0050] In some cases, when the second connector valve is in the second position, the one or more reagent containers and the one or more more bioprocessing chambers are fluidically sealed from the sterilization chamber.

[0051] In another aspect, provided herein is a kit, comprising: (a) a sterilization box; (b) a first connector valve comprising a first fluid line embedded therein, wherein the first connector valve is configured to releasably couple to the sterilization chamber such that the first connector valve forms a fluid tight seal against the sterilization chamber; and (c) a second connector valve comprising a second fluid line embedded therein, wherein the second connector valve is configured to releasably couple to the sterilization chamber such that the second connector valve forms a fluid tight seal against the sterilization chamber, and wherein the second connector valve is configured to move from a first position to a second position; wherein when the second connector valve is in the first position, the first connector valve and the second connector valve are closed such that the sterilization chamber is fluidically sealed from the first fluid line and the second fluid line, and wherein when the second connector valve is in the second position, the second connector valve engages with the first connector valve, thereby opening the first connector valve and the second connector valve such that the first fluid line and the second fluid line are fluidically connected. [0052] In another aspect, provided herein is a non-transitory computer readable medium comprising machine-executable code that, upon execution by one or more computer processors, implements a method for sterilizing a sterilization box, the method comprising: (a) providing a sterilization box comprising: (i) a sterilization chamber, (ii) a first connector valve comprising a first fluid line embedded therein, and (iii) a second connector valve comprising a second fluid line embedded therein, wherein the second connector valve is configured to move from a first position to a second position; (b) while the second connector valve is in the first position, introducing a sterilization fluid into the sterilization chamber, wherein when the connector valve is in the first position, the first connector valve and the second connector valve are closed such that the sterilization chamber is fluidically sealed from the first fluid channel and the second fluid channel, and the first connector valve and the second connector valve form a fluid tight seal against the sterilization chamber; and (c) adjusting a position of the second connector valve from the first position to the second position such that the second connector valve engages with the first connector valve, thereby resulting in the first fluid channel and the second fluid channel being fluidically connected, wherein the first fluid channel and the second fluid channel form a sterile fluid pathway.

[0053] In another aspect, provided herein is a sterilization apparatus, comprising: (a) a sterilization chamber configured to receive a sterilization fluid; (b) a first connector valve comprising a fluid line embedded therein, wherein said first connector valve forms a fluid tight seal against said sterilization chamber; and (c) a second connector valve, wherein said second connector valve forms a fluid tight seal against said sterilization chamber, wherein said second connector valve is configured to move from a first position to a second position, wherein when said second connector valve is in said first position, said first connector valve and said second connector valve are closed such that the sterilization chamber is fluidically sealed from said fluid line, and wherein when said second connector valve is in said second position, said second connector valve engages with said first connector valve, thereby opening the first connector valve such that said fluid line and said sterilization chamber are fluidically connected.

INCORPORATION BY REFERENCE

[0054] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and/or take precedence over any such contradictory material.

BRIEF DESCRIPTION OF THE DRAWINGS

[0055] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0056] FIG. 1 illustrates a schematic of system with three reagents and cassettes at a first latched position and prior to a sterilizing cycle, in accordance with some embodiments. [0057] FIG. 2 illustrates a schematic of system with three reagents and cassettes, at a first latched position after a sterilizing cycle, in accordance with some embodiments.

[0058] FIG. 3 illustrates a schematic of system with three reagents and cassettes, at a second latched position after a sterilizing cycle, in accordance with some embodiments.

[0059] FIG. 4 illustrates a schematic of a sterilizing box with three pairs of connector valves, in accordance with some embodiments.

[0060] FIG. 5 illustrates a cross-section view of a sterilizing box with a first connector in a latched position and a second connector in a first latched position, in accordance with some embodiments. [0061] FIG. 6 illustrates a cross-section view of a sterilizing box with a first connector in a latched position and a second connector in a second latched position, in accordance with some embodiments. [0062] FIG. 7 illustrates a first connector shown with its valve closed and a connector pin fully extended, in accordance with some embodiments.

[0063] FIG. 8 illustrates a cross-section view of a first connector shown with its valve in the open position with a fluid channel bridging a connector pin front seal, in accordance with some embodiments.

[0064] FIG. 9 schematically illustrates a computer system that is programmed or otherwise configured to implement methods provided herein.

DETAILED DESCRIPTION

[0065] While various embodiments have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions can occur to those skilled in the art without departing from the disclosure. It should be understood that various alternatives to the embodiments of the disclosure described herein can be employed.

[0066] Whenever the term “about,” “at least,” “greater than,” or “greater than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “about,” “at least,” “greater than” or “greater than or equal to” applies to each of the numerical values in that series of numerical values. For example, greater than or equal to 1, 2, or 3 is equivalent to greater than or equal to 1, greater than or equal to 2, or greater than or equal to 3.

[0067] Whenever the term “no more than,” “less than,” or “less than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “no more than,” “less than,” or “less than or equal to” applies to each of the numerical values in that series of numerical values. For example, less than or equal to 3, 2, or 1 is equivalent to less than or equal to 3, less than or equal to 2, or less than or equal to 1.

Overview

[0068] The devices, methods, and systems described herein are directed to a connector that operates in conjunction with a bespoke designed sterilizing box system that facilitates multiple sterilizing cycles for the connectors. This allows the devices described herein to be reused many times, including any connectors that attach to the reusable parts of the system. The sterilizing box and connector system described herein allow the external surfaces of the connectors that contact one another during fluid flow to be sterilized in a box that is sealed to the non-sterile environment prior to fluid connection with the mating connector. Some advantages of this reusable and re-sterilizable connector is that it reduces the consumable waste of the system when an experimental run is complete, reduces the cost associated with single-use connectors, and reduces setup time as the operator is not required to replace all the lines and connectors that require attachment.

Connection of Sterile Subsystems

[0069] The aseptic connectors described herein may be used to connect two or more separate and individual types of sterile subsystems. The sterile subsystems can be any microfluidic system designed to hold or transport reagents or biological components. For example, these sterile subsystems can include reagent tanks, valves, pumps, bioreactors, cassettes designed for bioprocessing, or waste tanks. In order for reagents and biological components to be transported from a first sterile system to a second sterile system, the sterile systems must be fluidically connected at least one point in time. The connectors and sterilization box described herein can be used at the interface of two or more sterile subsystems to fluidically connect the subsystems. [0070] For example, prior to starting an experiment, two or more separate and individual types of sterile subsystems may need to be brought into aseptic connection with each other. The types of subsystems can include, but are not limited to: one or more reagent tanks, a valve system with or without integrated pumps, one or more bioprocessing cassettes, and a waste vessel. Subsystems may be fluidically connected to each other through fluid lines. In some cases, each fluid line has a connector at the end of it which connects to a sterilizing box. After attachment, a gas tight seal can be formed between the connector and the sterilizing box. After connection of the subsystems to the sterilizing box and subsequent sterilization of the sterilization box, the entirety of the fluid system containing the reagents and cells can be enclosed within one single sterile barrier so that the culturing of the cells can be achieved in a closed-circuit sterile environment. A sterilization box can be used to fluidically connect two fluid lines. In some cases, a single sterilization box can be used to fluidically connect more than two fluid lines. For example, as shown in FIGs. 1-3, a single sterilizing box may contain many connections ports for all the connectors required by the system. In some cases, as shown in FIGs. 1-3, three sterilization boxes each with three connector ports allow three reagents and three bioprocessing cassettes all connect to one valve system. The number of sterilization boxes and ports required can be dependent on how many bioprocessing cassettes and reagents are chosen to be included in the system.

[0071] In FIGs. 1-3, individual sterile zones are shown by dashed boundaries. FIG. 1 illustrates a schematic of system with three reagents and three bioprocessing cassettes at a first latched position and prior to a sterilizing cycle, in accordance with some embodiments. In a first latched position, connectors can be used to attach input and output fluid lines to sterilization boxes 100. In this first latched position, the connectors can be attached to the sterilization boxes 100, thereby forming a gas tight seal against the sterilization box. In this first latched position, the valves in the connectors can be in a closed position such that there is no fluidic connection between the sterilization box and subsystems connected via the connector valves. The three reagent tanks 105 are each individually surrounded by a sterile boundary, resulting in a first, second, and third sterile boundary. The three bioprocessing cassettes 110 are each individually surrounded by a sterile boundary, resulting in a fourth, fifth, and sixth sterile boundary. A seventh sterile boundary exists around valve system 115 and waste output 120.

[0072] After the first latched position shown in FIG. 1 is achieved, the sterilizing box can undergo a sterilization cycle. A sterilization fluid can be fed into the sterilization box via a sterilization fluid input line. Because the connector valves can form a gas tight seal at the first latched position, sterilization fluid can remain in the sterilization box until it is removed via a sterilization fluid output line. A sterilization cycle can sterilize all internal surfaces of the sterilizing box, including the surfaces of the connectors that form the gas tight seal with the sterilization box. After a sterilization cycle, the sterilization box can become a sterile zone. FIG. 2 illustrates a schematic of system with three reagents and cassettes, at a first latched position after a sterilizing cycle, in accordance with some embodiments. As shown in FIG. 2, after a sterilization cycle, there are three additional sterile zones around each sterilization box, resulting in ten total individual sterile zones.

[0073] After sterilization, connectors can move to a second latched position. In this second latched position, pairs of connectors can engage with each other. For example, a first connector connected to a first fluid line can engage with a second connector connected to a second fluid line. The connectors can engage such that a gas tight seal is created between the two connectors. Additionally, the connectors can engage such that the valves in the connectors can move from a closed position to an open position, thereby resulting in the first fluid line and second fluid line being fluidically connected. Because the transition of the connectors can take place entirely within the sterile zone of the sterilization chamber, the sterile barrier is not broken. Therefore, as shown in FIG. 3, which illustrates the system at the second latched position after the sterilizing cycle, one sterile zone can be created that surrounds the entire system. Reagents and biological components can then be transported between the various subsystems without breaking sterility.

Sterilization Box and Aseptic Connectors

[0074] FIG. 4 illustrates a schematic of a sterilizing box with three pairs of connector valves, in accordance with some embodiments. Two unique types of connectors can be used to connect and seal fluid lines to the sterilizing boxes. A first connector can be designed to engage with its corresponding second connector to form a fluidic connection. A first connector may be referred to as a male connector. A second connector may be referred to as a female connector. Referring to FIG. 4, the connectors can be paired up such that every male connector (1) has a corresponding female connector (2), which it connects to after the sterilizing process is complete. A male connector may engage with a female connector such that at least a portion of the male connector is inserted into and surrounded by at least a portion of the female connector.

[0075] In some cases, a first connector can be inserted into a first side of the sterilization box. The corresponding second connector can be inserted into a second side of the sterilization box directly across from the first connector. Once connected to the sterilizing box, the connectors can become mechanically attached or locked to the sterilizing box. This may be referred to as a first latched position. In some cases, the connectors can form a fluid tight seal with the sterilization chamber such that fluid cannot escape the junction between the connector and the sterilization box. In some cases, the connectors are mechanically attached to the sterilization box via a spring-loaded latch (4). In some cases, the spring-loaded latch (4) must be depressed in order to release the connector.

[0076] Prior to commencing an experiment and connecting the male and female connectors together, the sterilizing boxes and the surfaces of the connectors that face into the sterilizing box may require sterilizing. To achieve this, a sterilizing fluid can be introduced into the sterilizing box. In some cases, the sterilization fluid is under pressure. In some cases, the sterilizing fluid is steam. The sterilizing fluid can be pressurized steam that is introduced to the sterilizing box at a pressure that relates to the temperature of the steam of at least 121 °C. In some cases, a steam generator creates the steam using a supply of water and heat. In some cases, the steam is saturated steam. In some cases, the steam is superheated steam. A steam distribution system can transport steam to each sterilizing box and the steam can enter via an inlet port 200. A sterilizing box can also have an exhaust port 205. The exhaust port can allow air in the sterilizing box to be displaced by the sterilizing fluid entering the sterilizing box such that the air exits out the sterilizing box and is transported into a steam condenser via the distribution system.

[0077] Using steam as the sterilizing fluid can offer many benefits. These benefits include: a short sterilizing cycle time, a working fluid of purified / distilled water that is readily available, low cost, non-toxic and non-corrosive. Materials such as stainless steels and high temperature plastics and elastomers including polysulfones, poly ether ether ketones (PEEK) silicones and fluorocarbon elastomers are mechanically robust to repeated steam sterilizing, and so these materials can be used in the construction of the sterilizing boxes, connectors, and steam system. Additionally, steam may have an additional benefit of being able to transport around a distribution system without the need for a pump. As steam is heated beyond boiling point its thermodynamic properties result in an increase in pressure compared to ambient atmospheric pressure. For instance, at 121 °C the gauge pressure of steam is approximately 1. IBar. This high pressure, relative to ambient, can be sufficient for steam to transport itself around a steam distribution system, displacing air that may be present in the process. Thus, a steam generator consisting of a boiler that delivers heat to the water, can be all that is required for steam transportation. Therefore, in some cases, other than valves to regulate the flow of the steam, no other moving parts are required. Another benefit of using steam is that steam under pressure is able to penetrate into narrow passageways and crevices.

[0078] Apart from steam, other sterilizing fluids can be used. The sterilization fluid can be a gas or a liquid. The sterilization fluid can be hydrogen peroxide liquid or vapour or ethylene oxide gas.

[0079] Both the male and female connectors can function as a valve that can either be in an open or closed state. The default position for both connectors is that their valves are closed. However, both connector’s valves can change to the open state when the male and female connectors connect to each other. In the closed state the connectors can maintain the sterile barrier of the fluid line that they are attached to. The closed valve can prevent microbes from the external environment crossing the valve and contaminating the fluid contained within the internally inside the connector.

[0080] The male and female connectors differ from one another in that the male connector (1) can have a single latched position with the sterilizing box whilst the female connector (2) can have two latched positions / states with the sterilizing box. For example, both the male and female can achieve a first latched position. This first latched position can be achieved when a connector is mechanically attached or locked to the sterilizing box. In this first latched position, a gas tight seal can formed between the outside diameters of the first and second connectors and the sterilizing box. In some cases, the male connector remains in this first latched position the entire time it is attached to a sterilization box.

[0081] FIG. 5 shows a cross-sectional view of a sterilizing box with a male connector in its first latched position and the female connector in its first latched position. When the female connector (2) is in its first latched position, the female connector can be attached and sealed to the sterilizing box but not fluidly connected to the male connector (1) or the sterilization chamber (3a). [0082] FIG. 6 shows a cross-sectional view of a sterilizing box with a male connector in its first latched position and the female connector in its second latched position. When the female connector (2) is in its second latched position, the female connector can be attached and sealed to the sterilizing box and fluidly connected to the male connector (1). In some cases, moving the position of the female connector from its first latched position to its second latched position results in two or more sterile subsystems becoming fluidically connected, thereby forming a single fluidically connected sterile system comprising two or more subsystems.

[0083] As shown in FIG. 5, during setup of an experiment, a user may first attach all the male connectors (1) to the sterilizing boxes, thereby resulting in the male connectors being in their first latched position. In some cases, all fluid lines (5) that lead to a sterile subsystem have a male connectors (1) that latches and seals to a sterilizing box. The user can then insert the female connectors (2) to the sterilizing boxes to their first latched position. In this state, the male and female connectors may not be fluidly connected to one another. When both the male and female connectors are in their first latched position, their valves can be in the closed state. After all the connectors are sealed to the sterilizing chamber (3a), the sterilized box is an enclosed sealed volume and can be sterilized by the introduction of a sterilization fluid (i.e. steam) from the sterilization fluid input (200). All internal surfaces exposed to the steam in the sterilizing box can be sterilized.

Additionally, the steam may sterilize the external faces of the connectors that come into contact with one another in step (c). These same surfaces can also be wetted by reagents once the connectors are fluidly connected and the experiment is underway.

[0084] As shown in FIG. 6, after the sterilizing cycle is complete, a user can reposition the female connectors (2) to their second latched position by pushing the female connectors (2) axially further into the sterilizing boxes. As the female connector (2) is pushed to the second latched position two things can happen in sequence. First, the outer shroud of the female connector (2) can make a gas tight seal with the outside diameter of the male connector (1). Second, by pushing the female connector (2) further into the sterilizing box, both the male and female connector’s valves can move to their open state. This can be achieved by connector pins (7) in each connector coming into contact with one another. When two connecting pins come in contact, they can be pushed into their respective connectors. This motion may cause the connector pins (7) to depress against their respective compression spring (lb). This motion may also open the fluid channels (7a) between the connectors, thereby allowing fluid connection between the female connector (2) and the male connector (1). Thus, after this step is complete the male and female connectors can be fluidly sealed to one another and both connector’s valves can be in the open state. Once this step has been achieved for all the connectors, all the separate systems with their respective sterile barriers can be brought into fluid connection in an aseptic manner such that when connection is complete only one sterile barrier is present that encompasses the entire fluid circuit, shown schematically in FIG. 3. [0085] Certain features present in the connectors can be used to achieve a valve function. The principle of the features for both male and female connectors may be the same. FIG. 7 and FIG. 8 schematically illustrate the features of the connector valves described herein, in accordance with some embodiments.

[0086] A connector can have a connector body which is hollow. In some cases, housed inside the body is a connector pin (7) which can be circular in form. In some cases, the connector pin (7) has a diameter smaller than the hole in the body to which it passes into. Seals can be placed at either end of the connector pin (7), which can result in a fluid tight radial seal (6) between the connector body and the connector pin (7). If the diameter of the connector pin (7) is smaller than the through hole diameter of the connector body, a space can exist between the two in a shape of an annulus.

[0087] A connector can contain a fluid line. The fluid line for the connector can enter the connector body at a certain angle relative to the axis of the connector body. The angle can be about 90°. In some cases, the angle is less than 90°. For example, the angle can be about 85°, 80°, 70°, or 45°. In some cases, the angle is greater than 90°. For example, the angle can be about 95°, 100°, 110°, or 135°

[0088] The fluid line can be attached to the connector body using either adhesive or through a barbed feature. FIG. 6 and FIG. 7 show the fluid line entering into a hole present in the connector body to which a small chamber at the entrance of the hole that can function as a well for adhesive to be applied to retain the fluid line to the connector body. The fluid line hole can connect to an annulus created between the connector pin (7) and the connector body. Therefore, fluid can occupy the internal volume within the connector. In some cases, the internal volume within the connector consists of the volume of the fluid line hole and the annulus volume, which can be sealed at either end of the annulus by the seals.

[0089] In some cases, the front of the connector pin (7) is located the end of the connector that is configured to be inserted into the sterilizing box (the proximal end). As such, in some cases, the connector pin (7) faces or points into the sterilizing box. The rearward or distal end of the connector pin (7) can have a shoulder (7b) which retains a compression spring (lb, 2b). The spring can be housed between the connector pin (7) and an end cap (9). The end cap can mechanically attach to the connector body. This mechanical attachment may comprise a bayonet, a screw thread, or an adhesive. In some cases, the end cap (9) is thermally bonded to the compression body by laser welding, ultrasonic welding or hot plate welding. In some cases, and as shown in FIGs. 5-9, the end cap has a bayonet channel which can engage a bayonet pin feature on the connector body.

[0090] When the connector is assembled, the compression spring (lb, 2b) can move from a preloaded state to a loaded state. In a preloaded state, the compression spring (lb, 2b) can apply a force between the end cap and the shoulder (7b) of the connector pin (7). The connector pin (7) can be pushed forward by the force of the compression spring (lb, 2b) until its shoulder comes into contact with an axial endstop. In some cases, the rearward connector pin seal (8b) acts as the endstop. The connector pin (7) may have a small fluid channel (7a) that extends axially along the pin from the front end of the connector pin (7). The fluid channel (7a) may be located at the proximal or front end of the pin.

[0091] When the male connector (1) is latched and the female connector (2) is in the second latched position, the connector pin (7) can be displaced into the connector body. When the connector pin (7) is displaced into the connector body, the fluid channel (7a) can allow fluids to flow through the fluid line (5) of the male connector, through the fluid channel (7a) of the male connector, through the fluid channel (7a) of the female connector, and through the fluid line (5) of the female connector. Fluid can flow either direction between the male and female. For example, fluid can flow from the male connector to the female connector or from the female connector to the male connector.

[0092] When the male connector (1) is latched and the female connector (2) is in the first latched position, the connector pin (7) can extend partially or fully forward relative to the connector body (i.e., the connector pin (7) is not displaced within the connector body). In this position, the entirety of the fluid channel (7a) can be located in front of the connector pin’s seals. Therefore, the fluid channel (7a) is not in fluid communication with the internal fluid volume of the connector, and so the state of the valve is closed. In this closed valve position, both the front and the rear connector pin seals (8) can make a complete circumferential seal between the connector body and the connector pin (7). Hence, the fluid inside the connector body can be enclosed and sealed within the internal volume of the connector body. In other words, the fluid inside the connector body cannot enter the sterilization chamber when the valve is in a closed position. Because the fluid channel (7a) is outside of the sealed volume of the connector body, it can exposed to a non-sterile environment. FIG. 7 shows a cross-sectional view the male connector in isolation but in the first latched position.

The fluid channel (7a) can protrude from the front connector pin seal (8), exposing it to a non-sterile environment. Therefore, to maintain sterility, the fluid channel (7a) can be sterilized before fluid flows through it.

[0093] When the male connector (1) is latched to the sterilizing box and the female connector (2) is latched to the first latched position, the radial seals between connectors and the sterilizing box (8) present on the outside diameter of the connectors can form a fluid tight seal to the sterilizing box. During the sterilizing cycle, steam or another sterilization fluid can come into contact with the end of the connector pin (7) that protrudes into the sterilizing chamber (3 a). In some cases, the fluid channel (7a) in the connector pin (7) protrudes into the sterilization chamber (3a) and can therefore be contacted with the steam or other sterilization fluid. Steam sterilizing or sterilization with another sterilization fluid can kill any microbes on the surfaces of the connectors in the sterilizing chamber (3a), including the fluid channels (7a), and the internal surfaces of the chamber. Therefore, by creating a seal to the sterilizing box and sterilizing, the end of the connector pin (7) and the fluid channel (7a) can return to a sterile condition after having been previously exposed to a non-sterile environment.

[0094] After a sterilizing cycle has been completed, the female connector (2) can be displaced axially into the sterilizing box to the second latched position. As this occurs, the end surface of the female connector’s pin, which faces into the sterilizing chamber (3a), can make contact with the end surface of the connector pin (7) in the male connector (1) that also faces into the sterilizing chamber (3a). In some cases, the male connector (1) is directly opposite the female connector (2). FIG. 6 shows a schematic of a sterilizing box (3) when the female connector is in its second latched position. In some cases, the connectors become sealed to one another prior to the connector pins (7) contacting one another. In this second latched position, both connector pins can be displaced into the body of their respective connector.

[0095] At the second latched position, the axial distance between the connector ends and/or end caps of the male and female connectors can be less than when the female connector (2) is in its first latched position. In some cases, this is due to the male connector (1) being fixed axially to the sterilizing box in its latched position, whilst the female connector (2) is pushed axially into the sterilizing box as it is moved from the first to the second latched position. Both connector pins (7) can have a fixed length and can be made from a rigid material. As the male and female connector (2) pins come in contact with one another, displacement of the female connector (2) to the second latched position can cause the gap between the distal end of the connector pins (7) and the end caps to reduce due to the motion of the female connector (2). Each connector pins can recede or retract into its respective hollow connector body when the female connector body is displaced to the second latched position. As the gap between the shoulders (7b) of the connector pins and the end caps is reduced, the compression springs (lb, 2b) can become further compressed.

[0096] The relative motion of the connector body to the connector pins (7) can result in the fluid channels (7a) bridging the front connector pin seals (8) in both connector bodies. If this happens, the valve state of both connectors can become open because as the fluid channels recede or retract into the connector body, they pass underneath the connector pin seal (8), which is shown in FIG. 7. [0097] Prior to the connector pin seals being bridged, axial motion of the female connector (2) can result in the female connector seal (la) present on the outside surface of the male connector (1) making a fluid tight seal with the inside diameter of a collar feature (2a) in the female connector (2). In some cases, the function of the female connector seal (la) is to fluidly seal the male and female connectors to one another thus preventing leakage of fluid into the sterilization box. Once this seal has been achieved, the fluid channels (7a) can bridge the front connector pin seals (8) in both connectors. Thus, a sealed fluid connection can be achieved between the two connectors. At this point fluid, can flow from the fluid line (5) leading to one connector, into the fluid line hole of that connector, around the annulus surrounding its connector pin (7), through the fluid channel (7a) of the connector, and into the fluid channel (7a) of the mated connector.

[0098] In some cases, system sterility is not compromised as all of the external surfaces of the male and female connector that are wetted when the connectors are mated together and fluid is flowing have been steam sterilized in the sterilizing box (3).

[0099] In some cases, the steps previously discussed can be reversed to separate any particular male and female pair of connectors, which for instance allow the removal of a reagent. There are a number of reasons why the operator may wish to disconnect the fluid connector but a common reason may be because a reagent bag or bottle has been depleted and requires replacing during an experimental run. As the steps to connect the connectors can be reversed to disconnect them, connecting and disconnecting connectors can be repeated on multiple occasions. Thus the connectors can be repeatedly sterilized and reused.

[0100] For example, a user may wish to remove a reagent bag and its associated female connector. The connector that attaches to the fluid line (5) of the reagent bag can be removed from the sterilizing box (3). The connector can be removed by depressing the corresponding latch and pulling the connector from the sterilizing box (3). As the female connector (2) is displaced away from the second latched position, the axial distance between the male and female connectors can increase. Due to the force exerted by the compression springs (lb, 2b), both the male and female connector pins can be displaced outwards relative to their respective connector bodies. At a predetermined placement, and prior to the female connector (2) seal losing contact with the collar in the female connector (2), the fluid channels (7a) in the connector pins (7) can no longer bridge the front seal of the connectors. Therefore, the valves in both connectors can close. Additional axial movement of the connector can then cause the female connector (2) seal to lose contact with the collar in the female connector (2). At this point, the male and female connectors are no longer sealed to one another. Further axial movement of the female connector (2) can eventually cause the female connector to be removed from the sterilizing box (3), thereby allowing removal of the reagent vessel, its fluid line (5), and connector.

[0101] The reagent bottle or bag may then be refilled aseptically, for instance under a laminar flow hood. Once the refill or replacement has been completed, the reagent, fluid line (5) and connector can once again be attached to the sterilizing box (3) following the connection steps previously described. Afterwards, the refilled reagent vessel can once again aseptically connect to the system. [0102] At the end of a cell culture or specific experiment, a user may need to clean the reusable parts of the system to remove the fluids in the hold-up volume and/or sterilize the fluid contacting surfaces so that the system can be stored in a sterile state. This can be achieved by inserting a dummy female connector into the sterilizing box (3) in lieu of a functional female connector. In some cases, a dummy connector has no internal moving parts. However, it can have a protruding shaft which displaces the male connector pin when inserted into the sterilizing box (3). Additionally, a dummy connector can have an external seal which seals the dummy connector to the sterilization box when it is inserted into a receiving port on the sterilization box. In some cases, the dummy connector does not have a collar feature (2a), so no fluid seal is made between the dummy connector and the male connector (1) when the dummy connector is inserted to the second latched position. Hence, the dummy connector can open the valve of the male connector when in the second latched position such that the male connector (1) is in fluid communication with the sterilizing chamber (3 a) of the sterilizing box (3).

[0103] To flush out the reusable parts of the system and to sterilize the surfaces that come into contact with fluid, pressurised steam (or another sterilization fluid) may be introduced to a valve system at the end of a cell culture. If all the female connectors, which are fitted to the disposable parts of the system, have been swapped for dummy connectors, pressurized steam (or another sterilization fluid) can flow from the valve system, down the fluid lines (5), and into the male connectors (1). As the male connector’s valves are in the open state, steam passes through the valve and into the sterilizing chamber (3a) which can initially be at a lower pressure and temperature than the steam. The flow of steam can flush out liquid from the valve system, reusable fluid lines (5), and reusable male connectors (1) into the sterilizing chamber (3a). Once inside the sterilizing chamber (3a), the displaced fluid and steam can exit the sterilization (3) via the outlet port 205 where it can be transported to a waste container via the steam distribution system.

[0104] At the end of the sterilizing cycle, the dummy connectors can be removed from the sterilizing box (3). The valves of the male (non-dummy) connectors can be shut before the dummy connector loses sealing contact with the sterilizing box (3) and is subsequently removed. This can result in the valve system, fluid lines (5), and the male connectors (1) being in a clean and sterile condition. In some cases, at this point, the closed male valves are ready to be used again.

[0105] The aseptic connector system described herein comprise one or more pairs of connectors. For example, a sterilization box may be configured to receive two, three, four, or five pairs of connectors. The amount of connectors needed can be based on the number of sterile subsystems that are connected.

Computer Systems

[0106] In an aspect, the present disclosure provides computer systems that are programmed or otherwise configured to implement methods of the disclosure, e.g., any of the subject methods for sterilization. Computer systems can be used to automate any method described herein. FIG. 9 shows a computer system 2001 that is programmed or otherwise configured to implement a method for fluid distribution. The computer system 2001 can be configured to, for example, automate or control the amount of sterilization fluid that enters a sterilization box, move a connector valve from a first position to a second position, or direct flow of a reagent or bioprocessing fluid from a first fluid line to a second fluid line. The computer system 2001 can be configured to adjust a flow rate or an amount of fluid flow into or through a fluid distribution device or system, based on one or more user inputs or sensor readings. The computer system 2001 can be further configured to adjust the flow rate or an amount of fluid flow into or through a fluid distribution device or system in order to optimize (i.e., increase or decrease) the volume of fluid or concentration of solids in each outlet stream. The computer system 2001 can be an electronic device of a user or a computer system that is remotely located with respect to the electronic device. The electronic device can be a mobile electronic device.

[0107] The computer system 2001 can include a central processing unit (CPU, also "processor" and "computer processor" herein) 2005, which can be a single core or multi core processor, or a plurality of processors for parallel processing. The computer system 2001 also includes memory or memory location 2010 (e.g., random-access memory, read-only memory, flash memory), electronic storage unit 2015 (e.g., hard disk), communication interface 2020 (e.g., network adapter) for communicating with one or more other systems, and peripheral devices 2025, such as cache, other memory, data storage and/or electronic display adapters. The memory 2010, storage unit 2015, interface 2020 and peripheral devices 2025 are in communication with the CPU 2005 through a communication bus (solid lines), such as a motherboard. The storage unit 2015 can be a data storage unit (or data repository) for storing data. The computer system 2001 can be operatively coupled to a computer network ("network") 2030 with the aid of the communication interface 2020. The network 2030 can be the Internet, an internet and/or extranet, or an intranet and/or extranet that is in communication with the Internet. The network 2030 in some cases is a telecommunication and/or data network. The network 2030 can include one or more computer servers, which can enable distributed computing, such as cloud computing. The network 2030, in some cases with the aid of the computer system 2001, can implement a peer-to-peer network, which can enable devices coupled to the computer system 2001 to behave as a client or a server. [0108] The CPU 2005 can execute a sequence of machine-readable instructions, which can be embodied in a program or software. The instructions can be stored in a memory location, such as the memory 2010. The instructions can be directed to the CPU 2005, which can subsequently program or otherwise configure the CPU 2005 to implement methods of the present disclosure. Examples of operations performed by the CPU 2005 can include fetch, decode, execute, and writeback.

[0109] The CPU 2005 can be part of a circuit, such as an integrated circuit. One or more other components of the system 2001 can be included in the circuit. In some cases, the circuit is an application specific integrated circuit (ASIC).

[0110] The storage unit 2015 can store files, such as drivers, libraries and saved programs. The storage unit 2015 can store user data, e.g., user preferences and user programs. The computer system 2001 in some cases can include one or more additional data storage units that are located external to the computer system 2001 (e.g., on a remote server that is in communication with the computer system 2001 through an intranet or the Internet).

[0111] The computer system 2001 can communicate with one or more remote computer systems through the network 2030. For instance, the computer system 2001 can communicate with a remote computer system of a user (e.g., an operator managing or monitoring the bioprocessing). Examples of remote computer systems include personal computers (e.g., portable PC), slate or tablet PC's (e.g., Apple® iPad, Samsung® Galaxy Tab), telephones, Smart phones (e.g., Apple® iPhone, Android- enabled device, Blackberry®), or personal digital assistants. The user can access the computer system 2001 via the network 2030.

[0112] Methods as described herein can be implemented by way of machine (e.g., computer processor) executable code stored on an electronic storage location of the computer system 2001, such as, for example, on the memory 2010 or electronic storage unit 2015. The machine executable or machine readable code can be provided in the form of software. During use, the code can be executed by the processor 2005. In some cases, the code can be retrieved from the storage unit 2015 and stored on the memory 2010 for ready access by the processor 2005. In some situations, the electronic storage unit 2015 can be precluded, and machine-executable instructions are stored on memory 2010.

[0113] The code can be pre-compiled and configured for use with a machine having a processor adapted to execute the code, or can be compiled during runtime. The code can be supplied in a programming language that can be selected to enable the code to execute in a pre-compiled or as- compiled fashion.

[0114] Aspects of the systems and methods provided herein, such as the computer system 2001, can be embodied in programming. Various aspects of the technology can be thought of as "products" or "articles of manufacture" typically in the form of machine (or processor) executable code and/or associated data that is carried on or embodied in a type of machine readable medium. Machineexecutable code can be stored on an electronic storage unit, such as memory (e.g., read-only memory, random-access memory, flash memory) or a hard disk. "Storage" type media can include any or all of the tangible memory of the computers, processors or the like, or associated modules thereof, such as various semiconductor memories, tape drives, disk drives and the like, which can provide non-transitory storage at any time for the software programming. All or portions of the software can at times be communicated through the Internet or various other telecommunication networks. Such communications, for example, can enable loading of the software from one computer or processor into another, for example, from a management server or host computer into the computer platform of an application server. Thus, another type of media that can bear the software elements includes optical, electrical and electromagnetic waves, such as used across physical interfaces between local devices, through wired and optical landline networks and over various airlinks. The physical elements that carry such waves, such as wired or wireless links, optical links or the like, also can be considered as media bearing the software. As used herein, unless restricted to non-transitory, tangible "storage" media, terms such as computer or machine "readable medium" refer to any medium that participates in providing instructions to a processor for execution.

[0115] Hence, a machine readable medium, such as computer-executable code, can take many forms, including but not limited to, a tangible storage medium, a carrier wave medium or physical transmission medium. Non-volatile storage media including, for example, optical or magnetic disks, or any storage devices in any computer(s) or the like, can be used to implement the databases, etc. shown in the drawings. Volatile storage media include dynamic memory, such as main memory of such a computer platform. Tangible transmission media include coaxial cables; copper wire and fiber optics, including the wires that comprise a bus within a computer system. Carrier-wave transmission media can take the form of electric or electromagnetic signals, or acoustic or light waves such as those generated during radio frequency (RF) and infrared (IR) data communications. Common forms of computer-readable media therefore include for example: a floppy disk, a flexible disk, hard disk, magnetic tape, any other magnetic medium, a CD-ROM, DVD or DVD-ROM, any other optical medium, punch cards paper tape, any other physical storage medium with patterns of holes, a RAM, a ROM, a PROM and EPROM, a FLASH-EPROM, any other memory chip or cartridge, a carrier wave transporting data or instructions, cables or links transporting such a carrier wave, or any other medium from which a computer can read programming code and/or data. Many of these forms of computer readable media can be involved in carrying one or more sequences of one or more instructions to a processor for execution.

[0116] The computer system 2001 can include or be in communication with an electronic display 2035 that comprises a user interface (UI) 2040 for providing, for example, a portal for an operator to monitor or track one or more steps or operations of the sterilization or fluid distribution methods and systems described herein. The portal can be provided through an application programming interface (API). A user or entity can also interact with various elements in the portal via the UI. Examples of UTs include, without limitation, a graphical user interface (GUI) and web-based user interface. [0117] Methods and systems of the present disclosure can be implemented by way of one or more algorithms. An algorithm can be implemented by way of software upon execution by the central processing unit 2005. For example, the algorithm can be configured to adjust a flow rate or an amount of fluid flow into or through a fluid distribution device or system, based on one or more sensor readings or user inputs. In some embodiments, the algorithm can be further configured can be configured to, for example, automate or control the amount of sterilization fluid that enters a sterilization box, move a connector valve from a first position to a second position, or direct flow of a reagent or bioprocessing fluid from a first fluid line to a second fluid line.

[0118] While preferred embodiments of the present disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. It is not intended that the disclosure be limited by the specific examples provided within the specification. While the disclosure has been described with reference to the aforementioned specification, the descriptions and illustrations of the embodiments herein are not meant to be construed in a limiting sense. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. Furthermore, it shall be understood that all aspects of the disclosure are not limited to the specific depictions, configurations or relative proportions set forth herein which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the disclosure described herein can be employed in practicing the disclosure. It is therefore contemplated that the disclosure shall also cover any such alternatives, modifications, variations or equivalents. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

CLAIMS WHAT IS CLAIMED IS:

1. A sterilization apparatus, comprising:

(a) a sterilization chamber,

(b) a first connector valve comprising a first fluid line embedded therein, wherein said first connector valve forms a fluid tight seal against said sterilization chamber; and

(c) a second connector valve comprising a second fluid line embedded therein, wherein said second connector valve forms a fluid tight seal against said sterilization chamber, wherein said second connector valve is configured to move from a first position to a second position, wherein when said second connector valve is in said first position, said first connector valve and said second connector valve are closed such that the sterilization chamber is fluidically sealed from said first fluid line and said second fluid line, and wherein when said second connector valve is in said second position, said second connector valve engages with said first connector valve, thereby opening the first connector valve and the second connector valve such that said first fluid line and said second fluid line are fluidically connected.

2. The sterilization apparatus of claim 1, wherein said first connector valve comprises a first hollow connector body with a first diameter, wherein said first hollow connector body is fluidically connected to said first fluid line, wherein said first connector valve comprises a first connector pin with a second diameter, and wherein said second diameter is greater than said first diameter.

3. The sterilization apparatus of claim 2, wherein said first connector pin is configured to move from an extended position to a retracted position, wherein when in said extended position, at least a portion of said first connector pin extends into said sterilization chamber, and wherein when in said retracted position, said first connector pin retracts into said first hollow connector body.

4. The sterilization apparatus of claim 3, wherein said at least portion of said first connector pin has a first fluid channel embedded therein.

5. The sterilization apparatus of claim 4, wherein when said first connector pin is in said extended position, said first fluid channel is located entirely within said sterilization chamber.

6. The sterilization apparatus of claim 4 or claim 5, wherein when said first connector pin is in said extended position, said first fluid channel is fluidically sealed from said first fluid line, thereby resulting in said first connector valve being closed.

7. The sterilization apparatus of any one of claims 4 - 6, wherein said first connector pin comprises a first front connector pin seal, and wherein when said first connector pin is in said extended position, said first front connector pin seal causes said first fluid channel to be fluidically sealed from said first fluid line, thereby resulting in said first connector valve being closed.

8. The sterilization apparatus of any one of claims 4 - 7, wherein when said first connector pin is in said retracted position, said first fluid channel is fluidically connected to said first fluid line, thereby resulting in said first connector valve being open.

9. The sterilization apparatus of any one of claims 4 - 8, wherein when said second connector valve is in said first position, said first connector pin is in said extended position.

10. The sterilization apparatus of any one of claims 4 - 9, wherein when said second connector valve is in said second position, said first connector pin is in said retracted position.

11. The sterilization apparatus of any one of claims 4 - 10, wherein said first connector valve further comprises a first compression spring, and wherein said first compression spring is configured to move from a first state to a second state, wherein said first compression spring is compressed more in said second state than said first state, wherein when said second connector valve is in said first position, said first compression spring is in said first state, and wherein when said second connector valve is in said second position, said first compression spring is in said second state.

12. The sterilization apparatus of claim 11, wherein said first connector pin comprises a first back connector pin seal configured to fluidically seal said first fluid line from said first compression spring.

13. The sterilization apparatus of any one of claims 4 - 12, wherein said second connector valve comprises a second hollow connector body with a first diameter, wherein said second hollow connector body is fluidically connected to said second fluid line, wherein said second connector valve comprises a second connector pin with a second diameter, and wherein said first diameter is greater than said second diameter.

14. The sterilization apparatus of claim 13, wherein said second connector pin is configured to move from an extended position to a retracted position, wherein when in said extended position, at least a portion of said second connector pin extends into said sterilization chamber, and wherein when in said retracted position, said second connector pin retracts into said second hollow connector body.

15. The sterilization apparatus of claim 14, wherein said at least portion of said second connector pin has a second fluid channel embedded therein.

16. The sterilization apparatus of claim 15, wherein when said second connector pin is in said extended position, said second fluid channel is located entirely within said sterilization chamber.

17. The sterilization apparatus of claim 15 or claim 16, wherein when said second connector pin is in said extended position, said second fluid channel is fluidically sealed from said second fluid line, thereby resulting in said second connector valve being closed.

18. The sterilization apparatus of any one of claims 15 - 17, wherein said second connector pin comprises a second front connector pin seal, and wherein when said second connector pin is in said extended position, said second front connector pin seal causes said second fluid channel to be fluidically sealed from said second fluid line, thereby resulting in said second connector valve being closed.

19. The sterilization apparatus of any one of claims 15 - 18 wherein when said second connector pin is in said retracted position, said second fluid channel is fluidically connected to said second fluid line, thereby resulting in said second connector valve being open.

20. The sterilization apparatus of any one of claims 15 - 19, wherein when said second connector valve is in said first position, said second connector pin is in said extended position.

21. The sterilization apparatus of any one of claims 15 - 20, wherein when said second connector valve is in said second position, said second connector pin is in said retracted position.

22. The sterilization apparatus of any one of claims 15 - 21, wherein said second connector valve further comprises a second compression spring, and wherein said second compression spring is configured to move from a first state to a second state, wherein said second compression spring is compressed more in said second state than said first state, wherein when said second connector valve is in said first position, said second compression spring is in said first state, and wherein when said second connector valve is in said second position, said second compression spring is in said second state.

23. The sterilization apparatus of claim 22, wherein said second connector pin comprises a second back connector pin seal configured to fluidically seal said second fluid line from said second compression spring.

24. The sterilization apparatus of any preceding claim, wherein said first fluid line is configured to fluidically connect to one or more reagent containers, and wherein said second fluid line is configured to fluidically connect to one or more bioprocessing chambers.

25. The sterilization apparatus of any one of claims 1 - 23, wherein said second fluid line is configured to fluidically connect to one or more reagent containers, and wherein said first fluid line is configured to fluidically connect to one or more bioprocessing chambers.

26. The sterilization apparatus of any preceding claim, wherein said sterilization chamber is configured to receive a sterilization fluid.

27. The sterilization apparatus of claim 26, wherein said sterilization fluid comprises steam.

28. The sterilization apparatus of claim 26 or claim 27, wherein said sterilization fluid is configured to sterilize interior surfaces of said sterilization chamber.

29. The sterilization apparatus of any one of claims 4 - 28, wherein a or the sterilization fluid is configured to sterilize said sterilization chamber and said first fluid channel when said first connector pin is in said extended position.

30. The sterilization apparatus of any preceding claim, wherein said sterilization box comprises one or more of said first connector valves and one or more of said second connector valves, and wherein each of said one or more first connector valves are paired with each of said one or more second connector valves.

31. The sterilization apparatus of any preceding claim, wherein said sterilization box further comprises a latch, wherein said latch is configured to move said second connector valve from said first position to said second position.

32. The sterilization apparatus of claim 31, wherein said latch is configured to be moved manually by one or more users.

33. The sterilization apparatus of claim 31 or claim 32, wherein

34. said latch is configured to be moved automatically by one or more mechanical elements.

35. The sterilization apparatus of any preceding claim, wherein when said second connector valve is in said second position, said second connector valve at least partially surrounds a portion of said first connector valve.

36. The sterilization apparatus of any preceding claim, wherein said first connector valve comprises a male connector.

37. The sterilization apparatus of any preceding claim, wherein said second connector valve comprises a female connector.

38. The sterilization apparatus of any preceding claim, wherein when said second connector valve is in said second position, said first fluid line and said second fluid line are fluidically sealed from said sterilization chamber.

39. A method, comprising:

(a) providing a sterilization box comprising: (i) a sterilization chamber, (ii) a first connector valve comprising a first fluid line embedded therein, and (iii) a second connector valve comprising a second fluid line embedded therein, wherein said second connector valve is configured to move from a first position to a second position,

(b) while said second connector valve is in said first position, introducing a sterilization fluid into said sterilization chamber, wherein when said connector valve is in said first position, said first connector valve and said second connector valve are closed such that the sterilization chamber is fluidically sealed from said first fluid channel and said second fluid channel, and said first connector valve and said second connector valve form a fluid tight seal against said sterilization chamber; and (c) adjusting a position of said second connector valve from said first position to said second position such that said second connector valve engages with said first connector valve, thereby resulting in said first fluid channel and said second fluid channel being fluidically connected, wherein said first fluid channel and said second fluid channel form a sterile fluid pathway.

40. The method of claim 38, wherein said first connector valve comprises a first hollow connector body with a first diameter, wherein said first hollow connector body is fluidically connected to said first fluid line, wherein said first connector valve comprises a first connector pin with a second diameter, and wherein said second diameter is greater than said first diameter.

41. The method of claim 39, wherein said first connector pin is configured to move from an extended position to a retracted position, wherein when in said extended position, at least a portion of said first connector pin extends into said sterilization chamber, and wherein when in said retracted position, said first connector pin retracts into said first hollow connector body.

42. The method of claim 40, wherein said at least portion of said first connector pin has a first fluid channel embedded therein.

43. The method of claim 41, wherein when said first connector pin is in said extended position, said first fluid channel is located entirely within said sterilization chamber.

44. The method of claim 41 or claim 42, wherein when said first connector pin is in said extended position, said first fluid channel is fluidically sealed from said first fluid line, thereby resulting in said first connector valve being closed.

45. The method of any one of claims 41 - 43, wherein said first connector pin comprises a first front connector pin seal, and wherein when said first connector pin is in said extended position, said first front connector pin seal causes said first fluid channel to be fluidically sealed from said first fluid line, thereby resulting in said first connector valve being closed.

46. The method of any one of claims 41 - 44, wherein when said first connector pin is in said retracted position, said first fluid channel is fluidically connected to said first fluid line, thereby resulting in said first connector valve being open.

47. The method of any one of claims 41 - 45, wherein when said second connector valve is in said first position, said first connector pin is in said extended position.

48. The method of any one of claims 41 - 46, wherein when said second connector valve is in said second position, said first connector pin is in said retracted position.

49. The method of any one of claims 41 - 47, wherein said first connector valve further comprises a first compression spring, and wherein said first compression spring is configured to move from a first state to a second state, wherein said first compression spring is compressed more in said second state than said first state, wherein when said second connector valve is in said first position, said first compression spring is in said first state, and wherein when said second connector valve is in said second position, said first compression spring is in said second state.

50. The method of claim 48, wherein said first connector pin comprises a first back connector pin seal configured to fluidically seal said first fluid line from said first compression spring.

51. The method of any one of claims 41 - 49, wherein said second connector valve comprises a second hollow connector body with a first diameter, wherein said second hollow connector body is fluidically connected to said second fluid line, wherein said second connector valve comprises a second connector pin with a second diameter, and wherein said first diameter is greater than said second diameter.

52. The method of claim 50, wherein said second connector pin is configured to move from an extended position to a retracted position, wherein when in said extended position, at least a portion of said second connector pin extends into said sterilization chamber, and wherein when in said retracted position, said second connector pin retracts into said second hollow connector body.

53. The method of claim 51, wherein said at least portion of said second connector pin has a second fluid channel embedded therein.

54. The method of claim 52, wherein when said second connector pin is in said extended position, said second fluid channel is located entirely within said sterilization chamber.

55. The method of claim 52 or claim 53, wherein when said second connector pin is in said extended position, said second fluid channel is fluidically sealed from said second fluid line, thereby resulting in said second connector valve being closed.

56. The method of any one of claims 52 - 54, wherein said second connector pin comprises a second front connector pin seal, and wherein when said second connector pin is in said extended position, said second front connector pin seal causes said second fluid channel to be fluidically sealed from said second fluid line, thereby resulting in said second connector valve being closed.

57. The method of any one of claims 52 - 55, wherein when said second connector pin is in said retracted position, said second fluid channel is fluidically connected to said second fluid line, thereby resulting in said second connector valve being open.

58. The method of any one of claims 52 - 56, wherein when said second connector valve is in said first position, said second connector pin is in said extended position.

59. The method of any one of claims 52 - 57, wherein when said second connector valve is in said second position, said second connector pin is in said retracted position.

60. The method of any one of claims 52 - 58, wherein said second connector valve further comprises a second compression spring, and wherein said second compression spring is configured to move from a first state to a second state, wherein said second compression spring is compressed more in said second state than said first state, wherein when said second connector valve is in said first position, said second compression spring is in said first state, and wherein when said second connector valve is in said second position, said second compression spring is in said second state.

61. The method of claim 59, wherein said second connector pin comprises a second back connector pin seal configured to fluidically seal said second fluid line from said second compression spring.

62. The method of any one of claims 38 - 60, wherein said first fluid line is configured to fluidically connect to one or more reagent containers, and wherein said second fluid line is configured to fluidically connect to one or more bioprocessing chambers.

63. The method of any one of claims 38 - 60, wherein said second fluid line is configured to fluidically connect to one or more reagent containers, and wherein said first fluid line is configured to fluidically connect to one or more bioprocessing chambers.

64. The method of any one of claims 38 - 62, wherein said sterilization fluid comprises steam.

65. The method of any one of claims 38 - 63, wherein said sterilization fluid is configured to sterilize interior surfaces of said sterilization chamber.

66. The method of any one of claims 41 - 64, wherein said sterilization fluid is configured to sterilize said sterilization chamber and said first fluid channel when said first connector pin is in said extended position.

67. The method of any one of claims 52 - 65, wherein said sterilization fluid is configured to sterilize said sterilization chamber and said second fluid channel when said second connector pin is in said extended position.

68. The method of any one of claims 38 - 66, wherein said sterilization box comprises one or more of said first connector valves and one or more of said second connector valves, and wherein each of said one or more first connector valves are paired with each of said one or more second connector valves.

69. The method of any one of claims 38 - 67, wherein said sterilization box further comprises a latch, wherein said latch is configured to perform said adjusting in (c).

70. The method of claim 68, wherein said latch is configured to be adjusted manually by one or more users.

71. The method of claim 68 or claim 69, wherein said latch is configured to be adjusted automatically by one or more mechanical elements.

72. The method of any one of claims 38 - 70, wherein when said second connector valve is in said second position, said second connector valve at least partially surrounds a portion of said first connector valve.

73. The method of any one of claims 38 - 71, wherein said first connector valve comprises a male connector.

74. The method of any one of claims 38 - 72, wherein said second connector valve comprises a female connector.

75. The method of any one of claims 38 - 73, wherein when said second connector valve is in said second position, said first fluid line and said second fluid line are fluidically sealed from said sterilization chamber.

76. A microfluidic system, comprising:

(a) one or more reagent containers;

(b) one or more bioprocessing chambers; and

(c) a sterilization apparatus comprising (i) a sterilization chamber, (ii) a first connector valve fluidically connected to said one or more reagent containers, wherein said first connector valve forms a fluid tight seal against said sterilization chamber, and (iii) a second connector fluidically connected to said one or more bioprocessing chambers, wherein said second connector valve forms a fluid tight seal against said sterilization chamber; wherein said first connector valve or said second connector valve is configured to move from a first position to a second position, wherein when said first connector valve or said second connector valve is in said first position, said first connector valve and said second connector valve are closed such that the sterilization chamber is fluidically sealed from said one or more reagent containers and said one or more bioprocessing chambers, and wherein when said first connector valve or said second connector valve is in said second position, said second connector valve engages with said first connector valve, thereby opening the first connector valve and the second connector valve such that said one or more reagent containers and said one or more bioprocessing chambers are fluidically connected.

77. The microfluidic system of claim 75, wherein said first connector valve comprises a first hollow connector body with a first diameter, wherein said first hollow connector body is fluidically connected to said first fluid line, wherein said first connector valve comprises a first connector pin with a second diameter, and wherein said second diameter is greater than said first diameter.

78. The microfluidic system of claim 76, wherein said first connector pin is configured to move from an extended position to a retracted position, wherein when in said extended position, at least a portion of said first connector pin extends into said sterilization chamber, and wherein when in said retracted position, said first connector pin retracts into said first hollow connector body.

79. The microfluidic system of claim 77, wherein said at least portion of said first connector pin has a first fluid channel embedded therein.

80. The microfluidic system of claim 78, wherein when said first connector pin is in said extended position, said first fluid channel is located entirely within said sterilization chamber.

81. The microfluidic system of claim 78 or claim 79, wherein when said first connector pin is in said extended position, said first fluid channel is fluidically sealed from said first fluid line, thereby resulting in said first connector valve being closed.

82. The microfluidic system of any one of claims 78 - 80, wherein said first connector pin comprises a first front connector pin seal, and wherein when said first connector pin is in said extended position, said first front connector pin seal causes said first fluid channel to be fluidically sealed from said first fluid line, thereby resulting in said first connector valve being closed.

83. The microfluidic system of any one of claims 78 - 81, wherein when said first connector pin is in said retracted position, said first fluid channel is fluidically connected to said first fluid line, thereby resulting in said first connector valve being open.

84. The microfluidic system of any one of claims 78 - 82, wherein when said second connector valve is in said first position, said first connector pin is in said extended position.

85. The microfluidic system of any one of claims 78 - 83, wherein when said second connector valve is in said second position, said first connector pin is in said retracted position.

86. The microfluidic system of any one of claims 78 - 84, wherein said first connector valve further comprises a first compression spring, and wherein said first compression spring is configured to move from a first state to a second state, wherein said first compression spring is compressed more in said second state than said first state, wherein when said second connector valve is in said first position, said first compression spring is in said first state, and wherein when said second connector valve is in said second position, said first compression spring is in said second state.

87. The microfluidic system of claim 85, wherein said first connector pin comprises a first back connector pin seal configured to fluidically seal said first fluid line from said first compression spring.

88. The microfluidic system of any one of claims 78 - 86, wherein said second connector valve comprises a second hollow connector body with a first diameter, wherein said second hollow connector body is fluidically connected to said second fluid line , wherein said second connector valve comprises a second connector pin with a second diameter, and wherein said first diameter is greater than said second diameter.

89. The microfluidic system of claim 87, wherein said second connector pin is configured to move from an extended position to a retracted position, wherein when in said extended position, at least a portion of said second connector pin extends into said sterilization chamber, and wherein when in said retracted position, said second connector pin retracts into said second hollow connector body.

90. The microfluidic system of claim 88, wherein said at least portion of said second connector pin has a second fluid channel embedded therein.

91. The microfluidic system of claim 89, wherein when said second connector pin is in said extended position, said second fluid channel is located entirely within said sterilization chamber.

92. The microfluidic system of claim 89 or claim 90, wherein when said second connector pin is in said extended position, said second fluid channel is fluidically sealed from said second fluid line, thereby resulting in said second connector valve being closed.

93. The microfluidic system of any one of claims 89 - 91, wherein said second connector pin comprises a second front connector pin seal, and wherein when said second connector pin is in said extended position, said second front connector pin seal causes said second fluid channel to be fluidically sealed from said second fluid line, thereby resulting in said second connector valve being closed.

-SO-

94. The microfluidic system of any one of claims 89 - 92, wherein when said second connector pin is in said retracted position, said second fluid channel is fluidically connected to said second fluid line, thereby resulting in said second connector valve being open.

95. The microfluidic system of any one of claims 89 - 93, wherein when said second connector valve is in said first position, said second connector pin is in said extended position.

96. The microfluidic system of any one of claims 89 - 94, wherein when said second connector valve is in said second position, said second connector pin is in said retracted position.

97. The microfluidic system of any one of claims 89 - 95, wherein said second connector valve further comprises a second compression spring, and wherein said second compression spring is configured to move from a first state to a second state, wherein said second compression spring is compressed more in said second state than said first state, wherein when said second connector valve is in said first position, said second compression spring is in said first state, and wherein when said second connector valve is in said second position, said second compression spring is in said second state.

98. The microfluidic system of claim 96, wherein said second connector pin comprises a second back connector pin seal configured to fluidically seal said second fluid line from said second compression spring.

99. The microfluidic system of any one of claims 75 - 97, wherein said sterilization chamber is configured to receive a sterilization fluid.

100. The microfluidic system of claim 98, wherein said sterilization fluid comprises steam.

101. The microfluidic system of claim 98 or claim 99, wherein said sterilization fluid is configured to sterilize interior surfaces of said sterilization chamber.

102. The microfluidic system of any one of claims 78 - 100, wherein said sterilization fluid is configured to sterilize said sterilization chamber and said first fluid channel when said first connector pin is in said extended position.

103. The microfluidic system of any one of claims 89 - 101, wherein said sterilization fluid is configured to sterilize said sterilization chamber and said second fluid channel when said second connector pin is in said extended position.

104. The microfluidic system of any one of claims 75 - 102, wherein said sterilization box comprises one or more of said first connector valves and one or more of said second connector valves, and wherein each of said one or more first connector valves are paired with each of said one or more second connector valves.

105. The microfluidic system of any one of claims 75 - 103, wherein said sterilization box further comprises a latch, wherein said latch is configured to move said second connector valve from said first position to said second position.

106. The microfluidic system of claim 104, wherein said latch is configured to be adjusted manually by one or more users.

107. The microfluidic system of claim 104 or claim 105, wherein said latch is configured to be adjusted automatically by one or more mechanical elements.

108. The microfluidic system of any one of claims 75 - 106, wherein when said second connector valve is in said second position, said second connector valve at least partially surrounds a portion of said first connector valve.

109. The microfluidic system of any one of claims 75 - 107, wherein said first connector valve comprises a male connector.

110. The microfluidic system of any one of claims 75 - 108, wherein said second connector valve comprises a female connector.

111. The microfluidic system of any one of claims 75 - 109, wherein when said second connector valve is in said second position, said one or more reagent containers and said one or more bioprocessing chambers are fluidically sealed from said sterilization chamber.

112. A kit, comprising:

(a) a sterilization box;

(b) a first connector valve comprising a first fluid line embedded therein, wherein said first connector valve is configured to releasably couple to said sterilization chamber such that said first connector valve forms a fluid tight seal against said sterilization chamber; and

(c) a second connector valve comprising a second fluid line embedded therein, wherein said second connector valve is configured to releasably couple to said sterilization chamber such that said second connector valve forms a fluid tight seal against said sterilization chamber, and wherein said second connector valve is configured to move from a first position to a second position; wherein when said second connector valve is in said first position, said first connector valve and said second connector valve are closed such that the sterilization chamber is fluidically sealed from said first fluid line and said second fluid line, and wherein when said second connector valve is in said second position, said second connector valve engages with said first connector valve, thereby opening the first connector valve and the second connector valve such that said first fluid line and said second fluid line are fluidically connected.

113. A non-transitory computer readable medium comprising machine-executable code that, upon execution by one or more computer processors, implements a method for sterilizing a sterilization box, the method comprising: (a) providing a sterilization box comprising: (i) a sterilization chamber, (ii) a first connector valve comprising a first fluid line embedded therein, and (iii) a second connector valve comprising a second fluid line embedded therein, wherein said second connector valve is configured to move from a first position to a second position;

(b) while said second connector valve is in said first position, introducing a sterilization fluid into said sterilization chamber, wherein when said connector valve is in said first position, said first connector valve and said second connector valve are closed such that the sterilization chamber is fluidically sealed from said first fluid channel and said second fluid channel, and said first connector valve and said second connector valve form a fluid tight seal against said sterilization chamber; and

(c) adjusting a position of said second connector valve from said first position to said second position such that said second connector valve engages with said first connector valve, thereby resulting in said first fluid channel and said second fluid channel being fluidically connected, wherein said first fluid channel and said second fluid channel form a sterile fluid pathway.

114. A sterilization apparatus, comprising:

(a) a sterilization chamber configured to receive a sterilization fluid;

(b) a first connector valve comprising a fluid line embedded therein, wherein said first connector valve forms a fluid tight seal against said sterilization chamber; and

(c) a second connector valve, wherein said second connector valve forms a fluid tight seal against said sterilization chamber, wherein said second connector valve is configured to move from a first position to a second position, wherein when said second connector valve is in said first position, said first connector valve and said second connector valve are closed such that the sterilization chamber is fluidically sealed from said fluid line, and wherein when said second connector valve is in said second position, said second connector valve engages with said first connector valve, thereby opening the first connector valve such that said fluid line and said sterilization chamber are fluidically connected.