WO2017132637A2

WO2017132637A2 - Apparatus and method for monitoring multiple fluidic samples

Info

Publication number: WO2017132637A2
Application number: PCT/US2017/015528
Authority: WO
Inventors: David Evans; Cory Gerdts
Original assignee: Protein Biosolutons
Priority date: 2016-01-29
Filing date: 2017-01-29
Publication date: 2017-08-03
Also published as: WO2017132637A3

Abstract

The present invention concerns an apparatus and method for simultaneously monitoring multiple fluid samples. The invention is specifically adapted to monitor an array of micro-fluidic conduits of a protein purification/crystallization device.

Description

APPARATUS AND METHOD FOR MONITORING MULTIPLE FLUIDIC

SAMPLES

Cross Reference to Related Application

This International application claims priority from U.S. Provisional Patent Application 62/288,874.

BACKGROUND OF THE INVENTION

FIELD OF THE INVENTION

The present invention concerns an apparatus and method for simultaneously monitoring multiple fluid samples. The invention is specifically adapted to monitor an array of micro-fluidic conduits of a protein purification/crystallization device. The system is designed for use with the protein purification device described in U.S. Patent 6,818,060, which is herein incorporated by reference, but may be adapted for use with other protein handling systems or any system having an array of fluid conduits, where monitoring the content of the conduits is desirable.

2. DESCRIPTION OF THE PRIOR ART

Macromolecular x-ray crystallography is an essential aspect of modern drug discovery and molecular biology. Using x-ray crystallographic techniques, the three-dimensional structures of biological macromolecules, such as proteins, nucleic acids, and their various complexes, can be determined at practically atomic level resolution. The enormous value of three-dimensional information has led to a growing demand for innovative products in the area of protein crystal lization, which is currently the major rate limiting step in x-ray structure determination.

One of the first and most important steps of the x-ray crystal structure determination of a target macromolecule is to grow large, well diffracting crystals with the macromolecule. As techniques for collecting and analyzing x-ray diffraction data have become more rapid and automated, crystal growth has become a rate limiting step in the structure

determination process.

Vapor diffusion is the most widely used technique for crystallization in modern macromolecular x-ray crystallography. In this technique, a small volume of the macromolecule sample is mixed with an approximately equal volume of a crystallization solution. The resulting drop of liquid (containing macromolecule and dilute

crystallization solution) is sealed in a chamber with a much larger reservoir volume of the crystallization solution. The drop is kept separate from the reservoir, either by hanging from a glass cover slip or by sitting on a tiny pedestal. Over time, the crystallization drop and the reservoir solutions equilibrate via vapor diffusion of the volatile species.

Supersaturating concentrations of the macromolecule are achieved, resulting in crystallization in the drop when the appropriate reservoir solution is used.

The process of growing biological macromo!ecuie crystals remains, however, a highly empirical process. Macromolecuiar crystallization is a hyperdimensional phenomena, dependent on a host of experimental parameters including pH, temperature, and the concentration of salts, macromoleeules, and the particular precipitating agent (of which there are hundreds). A sampling of this hyperspace, via thousands of crystallization trials, eventually leads to the precise conditions for crystal growth. Thus, the ability to rapidly and easily generate many crystallization trials is important in determining the right conditions for crystallization. Also, since so many multidimensional data points are generated in these crystallization trials, it is imperative that the experimenter be able to accurately record and analyze the data so that promising conditions are pursued, while no further time, resources, and effort are spent on negative conditions.

Recently, an international protein structure initiative has taken shape with the goal of determining the three dimensional structures of all representative protein folds. This massive undertaking in structural biology which may some day rival the human genome sequencing project in size and scope, is estimated to require a minimum of 100,000 x-ray structure determinations of newly discovered proteins for which no staictural information is currently available or predicted. For perspective, the total number of reported novel crystal structures determined to date (spanning nearly 50 years of work) is only approximately 10,000.

Using existing methods for the crystallization of proteins (random screens of conditions), the protein structure initiative will require a minimum of approximately 100 million crystallization trials. In addition, the biological information gleaned from genomic research in the protein structure initiative are expected to create even more demand for structural information. Specifically, the biotechnology and pharmaceutical industries are estimated to require upwards of tenfold more protein crystallization experiments (one billion) as a result of research and stmcture based dmg design and the use of crystallized therapeutic proteins. This would require that each of the approximately 500

macromolecular crystallography labs worldwide be responsible for setting up

approximately 2000 crystallization trials every working day of the year for five years. Currently, there is no known device available for setting up analysis macromolecular crystallization data on this scale.

The protein purification/crystallization device described in U.S. Patent 6,818,060 allows for conducting multiple crystallization trial simultaneously by providing screening fluid through an array of precisely metered fluid pumps connected to an array of micro-fluidic conduits.

Herein, we describe a novel apparatus and method for parallel UV monitoring of an array of fluid handling conduits, the system designed especially for, but not limited to, micro- fluidic lines.

SUMMARY OF THE INVENTION

It is a major object of the invention to provide a novel apparatus and method for parallel UV monitoring of fluidic lines. These and other objects of the present invention will become readily apparent upon further review of the following specification and drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

Various other objects, features, and attendant advantages of the present invention will become more fully appreciated as the same becomes better understood when considered with the accompanying drawings, in which like reference characters designate the same or similar parts throughout the several views, and wherein:

FIG. 1 is a schematic diagram illustrating a protein purification device to which the UV monitoring apparatus of the invention is attached.

FIG. 2 is a schematic diagram illustrating the generation and use of matrix recipes with the protein purification device of FIG. 1.

FIG. 3 is an illustration of an embodiment of the matrix maker of FIG. 2.

FIG. 4 is an illustration showing an array of stock solution containers and the tubing through which the solution passes, as employed in the embodiment of FIG. 3.

FIG. 5 is an illustration showing the pumps in the embodiment of FIG. 3. FIG. 6A is an illustration showing, in the embodiment of FIG. 3, the outlet manifold mounted to a gantry.

FIG. 6B is an illustration, similar to FIG. 6A, showing the gantry in a different position.

FIG. 7 is a closeup illustration of the embodiment of FIG. 3, showing the dispensing pins sticking through the outlet manifold.

FIG. 8 is a schematic diagram illustrating the operation of an embodiment of the invention, called the "Protein Maker-Drop Maker Robot."

FIG. 9 is an illustration showing, in the embodiment of FIG. 8, the outlet manifold mounted to a gantry.

FIG. 10 shows a front view of the UV monitoring system of the invention positioned on the gantry.

FIG. 11 shows a perspective view of FIG. 10.

FIG. 12 shows a perspective view of the system of FIG. 10 detailing the detector array. FIG. 13 shows a side perspective view of the system of FIG. 10 detailing the detector array.

FIG. 14A shows a sectional view detailing an individual detector.

FIG. 14B shows a perspective view detailing an individual detector.

FIG. 15 shows a perspective view, partly in section, detailing a row of detectors.

FIG. 16 shows a perspective view of the system housing illustrating the positioning of the electrical and optical systems.

FIG. 17 shows a detail of FIG. 16.

FIG. 18 shows a detail of the electrical and optical components. FIG. 19 shows a screen image from the software user interface of the invention. FIG. 20 shows a screen image from the software user interface of the invention. FIG. 21 shows a screen image from the software user interface of the invention. FIG. 22 shows a screen image from the software user interface of the invention. FIG. 23 shows a screen image from the software user interface of the invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

A description of preferred embodiments of the invention follows.

FIG. 1 is a schematic diagram illustrating the operation of an exemplary protein purification device to which the UV monitoring apparatus of the invention is attached.

The protein purification device uses positive pressure displacement of stock solutions through independently controlled precision syringe pumps, such that no disposable pipette tips are required. Viscous stock solutions can be delivered with high accuracy and speed through stainless steel outlet pins that do not come into contact with the recipient plasticware or reservoirs solutions as they are being created.

Proven "workhorse" precision syringe pumps, such as those manufactured by Tecan Systems (formerly Cavro Scientific Instruments, Inc.), can be used, minimizing subsequent maintenance.

Different sizes of syringes can be used to meet varying accuracy and scalability needs. More stock solutions can be added to the system as needed or demanded. Sterility of stock solutions can be maintained by eliminating open exposure to air. Finally, the appropriate volumes of stock solutions can be delivered directly to a crystallization plate or into sample tubes through the use of a manifold which is attached to a robotic gantry that can move in the X, Y and Z directions (e.g., lateral width, lateral depth and vertical directions). The outlet lines "shoot" stock solutions into the recipient plate (i.e., the individual solution receptacles).

Despite extensive investigation, the inventors were unable to identify a commercial liquid handling device that met these specifications.

Referring now to FIG. 1, plural (e.g., forty-eight) bottles 10 holding various stock solutions are directly connected via Teflon™ tubing 12 (of the shortest reasonable length) to the inlet ports of, for example twenty-four, individual precision syringe pumps 14, such as Tecan Systems' Cavro XL-3001. Each pump 14 is equipped with an 8-port distribution valve 16. In the illustrated embodiment, each pump 14 is attached through its distribution valve 16 to inlet lines 18 for two (2) different stock solutions, at valve positions 1 and 8.

Each pump 14 has two outlet lines 20, at valve positions 2 and 7, one for each of the two stock solutions. These outlet lines 20 are attached via tubing 44 (with the shortest reasonable distance) to an array of stainless steel dispensing pins or nozzles 26 held by an outlet manifold 28, which itself may be constructed from metal or some other suitable material.

Each pump 14 also has two waste outlet lines 22, for example at valve positions 3 and 6, through which waste is dumped into a waste container 42. In addition, each pump 14 has two water inlet lines 24, for example at valve positions 4 and 5, connected to a water/wash supply 40.

The outlet manifold 28 is mounted to a robotic gantry system 48 (see, for example, FIG. 6A) that can move the outlet manifold 28 in the X, Y, and Z directions/dimensions. The twenty-four pumps 14 are controlled by a controller 32. The robotic gantry 48 is controlled by a gantry controller 34. The pump/valve controller 32 and the gantry controller 34 shown in FIG. 1 may comprise both software components and hardware components (collectively, generally referred to as a processor system). The UV monitoring device of the present invention is attached to the robotic gantry 48, to receive fluid from, e.g., conduits 44 as will be explained in more detail later.

In one embodiment, the stock solution bottles 10, inlet lines 12, valves 16, and syringe pumps 14 are comprised of chemically resistant materials such as Teflon™,

polyetheretherketone (also known as PEEK™), glass, and stainless steel, such that the entire liquid path can withstand extreme pHs, high ionic strengths, and organic solvents. In this embodiment, the entire liquid path can be sterilized with chemical reagents such as ethanol, followed by extensive water washing and priming with filter sterilized stock solutions.

Solution receptacles, into which the various solutions are delivered, may be placed on a platform below the outlet manifold 28. For example, the solution receptacles may be wells arranged in an array on a crystallization plate 36, or tubes held in a tube rack 38. Multiple plates and/or tube racks may be positioned on the platform, and the software programmed accordingly to fill the containers of the various plates and tube racks.

In a preferred embodiment, the solution receptacles are stationary while the delivery system is positioned by the robotic gantry 48.

Software 30, such as Crystal Monitor™, available from Protein Biosystems, Inc., of Gaithersburg, MD (USA), provides for simple creation of a "recipe" for making a new set of screening solutions in the desired recipient crystallization plate 36 or rack 38 of tubes.

For example, the software 30 may have the ability to capture distance constraint information on plasticware and tube racks. The software can also calculate the volume of stock solutions needed to create a new crystallization screening matrix. It also has a knowledge base of the viscosity of each stock solution.

Database (DB) tables and graphical user interface (GUI) modules of software 30 may be used to perform the following: a) map stock solutions to physical positions on the invention; b) map stock solution inlet lines to valve positions on syringe pumps; c) map stock solution outlet lines to valve positions on syringe pumps; d) map stock solution outlet lines to outlet manifold pin positions on the gantry; and e) provide a knowledge base of titration curves for the final pHs achieved from mixing variable quantities of buffer stocks at 1 pH unit above and below the pKa of the buffer.

FIG. 2 is a schematic diagram illustrating the generation and use of the recipes. A user 101 may communicate with software 30 such as Crystal Monitor through a graphical user interface (GUI) driver 103, to define the system configuration as well as crystallization trial matrix solution specifications. A calculator 105 portion of the software 30 generates the "recipes", which may be stored, for example, in a table 107 in a database 109. Note that the system configuration information may also be stored in a table 108 in the same or a different database. The GUI driver 103 and calculator 105 may be integral parts of, for example, the matrix manager software described in U.S. application Ser. No. 09/631, 185.

Robot mixer control software 111 also contains a GUI driver 113, which may be launched by the Crystal Monitor program 30. The robot mixer control software 111 allows the user 101 to directly view and edit the contents of the recipe table 107.

Crystal Monitor 30 is able to launch the robot mixer driver 115 upon an appropriate user action. Based on the configurational information 108 and the recipes 107, the robot mixer driver 115 can generate a sequence of instructions or commands to control the matrix maker robot 200, by driving the syringe pumps 14, valves 16 (FIG. 1), and gantry 48 (see FIG. 3) in precise concert and sequence to deliver the appropriate stock solutions into the desired recipient containers.

The robot mixer driver 115 takes into account several considerations, including, but not limited to, the following:

(1) The final pH's for buffers are achieved by delivering the appropriate volumes of stock buffers which bracket the buffer pKa by pH unit (with reference to experimentally determined titration curves);

(2) The delivery sequence for different chemical types should be optimized;

(3) Travel distances should be minimized;

(4) Scheduling of pump wash cycles should be efficient; and

(5) "Chemical compatibility" features may be provided that warn the user that chemical precipitation would occur upon mixing certain chemicals (e.g., Ca2+ and phosphate are incompatible).

The table below illustrates a few exemplary rows (recipes) as might be defined in the recipe table 107. Although twelve columns are shown, it would be understood by one skilled in the art that other columns can be added for various purposes. However, only those columns needed to demonstrate the present invention are shown.

The "Dispensation No." is simply an identifier to identify a particular row in the table 107. Here, ten recipe rows are shown, having dispensation numbers from 58168 to 58177 respectively. The matrix mixer driver 115 may be capable of controlling multiple mixer robots 200. Therefore, the "Robot ID" column serves to identify the particular robot 200 to which the row pertains. Here, all rows pertain to robot #2.

A matrix may be given a name, specified in the "Matrix Name" column.

A robot may be capable of processing multiple trays simultaneously. Each tray maybe identified by a unique identifier. Here, all of the rows pertain to tray #12 of robot #2.

"Reagent No." specifies which stock solution bottle (10 from FIG. 1) is to be pumped, and the "Row" and "Col" columns specify the position of the receiving container that is to receive the identified stock solution. The "Vol" column indicates the volume to be dispensed, here in microliters.

So, for example, the first five rows, identified as dispensation numbers 58168 through 58172, direct that various stock solutions (from bottles numbered 19, 45, 11, 13 and 1 respectively) be dispensed into the container positioned at row 1, column 1, for tray 12 at robot 2, resulting in a 2-milliliter solution. The next five rows, identified as dispensation numbers 58173 through 58177 specify the solution to be mixed in the

receptacle/container at row 1, column 2 of the same tray.

The "Asp", "Disp", and "Drop" flags are simply flags used to indicate whether a respective particular operation has been done yet. For example, in the row for

dispensation no. 58170, the Asp flag (=Yes) indicates that aspiration has been performed, that is, the reagent from bottle 11 has been drawn into the corresponding pump 14. The Disp flag (=No) indicates that the stock solution has not yet been dispensed from the pump 14.

After dispensation, a drop of the stock solution may be hanging from the end of the dispensing pin. To prevent this drop from falling into and contaminating the dispensed solutions when the gantry is moved, an additional "drop" operation may be performed to draw back the drop (say about 5 microliters) into the dispensing pin. The "Drop" flag indicates whether this operation has been performed.

Finally, the status flag is used to indicate current status to the Crystal Monitor software 30 (FIG. 1).

Dispensation Robot Matrix Tray Reagent Asp Disp Drop Status

No ID Name TD No Row Col Vol Flag Flag Flag Flag

58168 2 matrixOOl 12 19 1 1 200 No No No 1

58169 2 matrixOOl 12 45 1 1 80 No No No 1

58170 2 matrixOOl 12 11 1 1 367.5373 Yes No No 1

58171 2 matrixOOl 12 13 1 1 32.46267 Yes Yes No 1

58172 2 matrixOOl 12 1 1 1 1320 Yes Yes Yes 1

58173 2 matrixOOl 12 19 1 2 200 Yes Yes Yes 1 58174 2 matrixOOl 12 45 1 2 80 Yes Yes Yes 1

58175 2 matrixOOl 12 11 1 2 312.7144 Yes Yes Yes 1

58176 2 matrixOOl 12 13 1 2 87.28554 Yes Yes Yes 1

58177 2 matrixOOl 12 1 1 2 1320 Yes Yes Yes 1

The matrix mixer driver 115 can accommodate various syringe sizes (e.g., 0.25 to 25 mL) and syringe speeds, different volume settings, etc.

FIG. 3 is an illustration of an embodiment of the matrix mixer 200 of FIG. 2. Stock solution bottles 10 are seated along either side of the platform 60. The stock solutions 10 are connected to the syringe pumps 14 via inlet tubing 12. The pumps 14 and their 8- position valves 16 sit atop a housing 50, which contains the gantry drive system for positioning the robotic gantry 48.

The outlet manifold 28 sits on the robotic gantry 48. Outlet tubing 44 connects the pumps 14 with the dispensing pins 26 which deliver the various solutions.

Here, the outlet manifold 28 has been positioned over a wash/waste receptacle 43 which sits on the platform or deck 60. The wash/waste receptacle 43 shown is of sufficient size (with respect to area) such that as many as all of the syringes and outlet tubes 44 may be washed simultaneously.

FIG. 4 is an illustration showing an array of stock solution containers 10 and the Teflon tubing 12 through which the solution passes, as employed in the embodiment of FIG. 3. FIG. 5 is an illustration showing the pumps 14 in the embodiment of FIG. 3. A first row of pumps 14 is located on top of the housing 50. A second row of pumps 14 is located behind the first row and is not visible in the FIG. 5 view. As can be seen from the figure, each pump 14 is attached to an associated 8-position valve 16 previously described in detail.

FIG. 6A is an illustration showing, in the embodiment of FIG. 3, the outlet manifold 28 mounted to the gantry 48. Tubing 44 from the pump valve outlets 20 (FIG. 1) is brought to the outlet manifold 28, and is connected to an array of dispensing pins 26. A wash/waste receptacle 42 is located on a stable platform next to a tube rack 38.

FIG. 6B is an illustration similar to FIG. 6A showing the gantry 48 in a different position with respect to tube rack 38 and wash/waste receptacle 42.

The tube rack 38 may be positioned to the platform/deck 60 via mounting pins (not shown) that allow the tube rack to be accurately positioned yet easily removed as an entire unit. This worktable mounting pin system provides the flexibility to utilize various racks containing different quantities of test tubes or different size test tubes, micro-plates, etc.

FIG. 7 is a closeup illustration of the embodiment of FIG. 3, showing the dispensing pins 26 sticking through the outlet manifold 28. Here, one solution 54 is being delivered to a receiving test tube 52, located in the test tube rack 38. It should be understood that multiple solutions may be delivered or dispensed to multiple receiving containers simultaneously.

In an alternate embodiment, several syringe pistons may be attached to a common drive, as for example, on the Cavro XL-3000-8. Thus, when one syringe piston is moving to deliver liquid, the other seven syringe pistons also move with the exact same stroke. However, the switch valves at the top end of each syringe are independently operated. Hence, when the XL-3000-8 performs a single liquid delivery cycle, the switch valve for the desired stock solution is the only one switched to the output position. The other stock solutions are pumped back into the stock bottles.

In this embodiment, the stock solutions can be arranged such that they are attached to the 8-position syringe drivers in an order that provides minimal chance that a given syringe pump would have to operate through more than one cycle during the construction of a single crystallization solution. For example, stock solutions which have similar chemicals may be attached to the same 8-port precision syringe pump. Then, following recipes of table 107, the matrix mixer driver 115 controls mixer robots 200 to pump/dispense through a subject syringe pump 14 once per cycle accordingly.

FIG. 8 is a schematic diagram illustrating the operation of another embodiment 800 of the invention, called a "Protein Maker-Drop Maker Robot." Solution inlet lines 801 are attached to an array of stainless steel pins or nozzles 26 held by a manifold 802. The manifold 802 is mounted to a robotic gantry system 48 (see, for example, FIG. 6A), which is controlled by software 804 via a gantry controller 34. The gantry controller 34 can control the movement of the manifold 802 in these orthogonal directions or dimensions. In this way, the pins 26 can be moved into sample plates 803 that contain desired solutions, which may be, for example, crude cell extracts containing protein, solutions containing purified protein, or chemical stock solutions. The pins, tubing and pumps involved are normally washed between aspiring different solutions to prevent contamination.

Alternatively, some pins could be offset from the rest and used individually without interference by the other pins.

Specified volumes of the solutions can be drawn into the inlet lines 801, by the appropriate specified valve 16 (see, for example, FIG. 1) and pump 14 movements under the control of software 804 via the pump controller 32. The solutions can be drawn into the syringe pumps 14, and then pumped through chromatography cartridges 807, via outlet lines 809, after the valves 16 change position to connect the pump 16 contents to the outlet lines 809.

The chromatography cartridges 807 are attached to an array of stainless steel dispensing pins or nozzles 26 held by the manifold 802. The solutions that flow through the chromatography cartridges 807 can be collected in collection plates 811, by software 804 controlled gantry movements of the manifold 802. Using specified pump 14 and valve 16 movements, the chromatography cartridges 807 can be washed with a plurality of different solutions (for example, wash buffer, equilibration buffer, elution buffers), which are attached to designated inlet valve 16 positions via additional inlet lines 813. The solutions that flow through the chromatography cartridges 807 can be collected in collection plates 811, by software 804 controlled gantry movements of the manifold 802.

Solutions from collection plates 811, protein sample plates 815, plates 817 containing detergents, plates 819 containing a set of ligands, and/or plates 821 containing

crystallization screening solutions prepared from stock solutions by, for example, the matrix maker 200, can be sequentially aspirated into solution inlet lines 801 and dispensed into crystallization plates 823 from the solution inlet lines 801 by the appropriate software 804 controlled pump 14, valve 16, and gantry 48 movements. The inlet lines 801 can be flushed with water between each aspiration and dispensing cycle. The water flush can be captured in the tip washer station 43 by the appropriate software 804 controlled pump 14, valve 16, and gantry 48 movements.

It should be apparent to one skilled in the art that the Matrix Maker Robot 200 and the Protein Maker-Drop Maker Robot 800 embodiments enable scientists to prepare new crystallization screening solutions from stock solutions, purify proteins from crude cell extracts, and set up crystallization plates by drawing from solutions in plates that were produced by the same embodiments.

FIG. 9 is a closeup illustration of the embodiment of FIG. 8, showing the dispensing pins 26 sticking through the outlet manifold 802. Here, twenty four chromatography cartridges 807 are mounted onto the manifold 802 and attached to outlet lines 809. Also shown are twenty-four inlet lines 801 that are attached to the manifold 802. The gantry 48 is shown directing the movement of inlet pins 26 into a sample plate 803. A collection plate 811 is also shown.

Referring now to Figs. 10 - 18, the fluid monitoring apparatus of the invention, generally indicated by the numeral 300, is shown. The apparatus 300 uses a source of UV light and a corresponding UV detector to determine the absorbance of a fluid flowing through a particular conduit. The UV light source and detector are positioned to measure the absorbance of the fluids coming off of the purification column. The apparatus 300 can be attached to gantry 48 by a suitable secure fastening means, this positioning allowing for receiving a plurality of conduits e.g., conduits 44. It can be seen that the apparatus 300 has a generally rectangular main housing 312, and can be adapted for use with any multiple throughput protein processing device having an array of fluid handling conduits for providing, e,g, screening fluid to a corresponding array of receptacles. It should be noted that attachment and positioning of the apparatus 300 of the present invention would be readily apparent to one of skill in the art. Also, the apparatus 300 of the present invention can be readily adapted for use in any system having multiple small diameter fluid conduits where monitoring of the fluid flowing through the conduits is desirable.

In order to monitor 24 different fluidic lines, 24 separate detection cells 302 are required, one for each line. In this design, the detection cells are grouped into 4 groups of six and arranged in a stair step format. Each detection cell 302 sends data that is recorded and displayed through the user interface 103. The stair step format is used to provide ample spacing for the fiber optic cables 304 to be organized and routed to the PC boards.

FIG. 13 is a view from the side of the stair-stepped detector arrays. Here you can see the connection of the fiber optic cables 304 that will bring in the required UV light. The black fittings 314 on top of the stair steps 316 are nuts for bringing in and securing tubing 317 that will be carrying the fluid to be analyzed.

FIG. 14A shows a cross-section of the detection cell 302 of the apparatus 300. The fiber optic cable 304 brings in the UV light from the light source 321. The detector 322 is opposite the fiber optic cable. The fluid enters the apparatus via tubing, which may be tubing 44, the tubing 44 secured in place at the top of the detection cell 302 by fastener 314 and ferrule 324. Just below the top ferrule 324 is a short piece of tubing 326 that serves as the flow cell. It is in this flow cell 326, which is preferably made of quartz, that the fluid is monitored as it moves through by an absorbance reading given by the relative difference in the reading provided by the detector 322 when the transmitted light reaches it.

FIG. 14B shows another cross-sectional view of the same region. Here, the light comes in from the left and is detected by the detector 322 on the left. The position of the flow cell 326 is between two ferrules. The bottom ferrule 328 holds a piece of tubing 329 in place used to route the fluid out of the apparatus 300.

FIG. 15 is yet another view of the detection region that includes an entire row of the detection cells 302 with the housing 312 made transparent. In this view, it can be seen how the detector leads 330 are routed back through the housing 312 to be connected to a PC board and allow for electrical connection to, e.g., the protein purification device described above or other processing and display means.

The electrical and optical components of the system are powered and controlled by 4 Arduino® controllers 340 as can be seen in Figs. 16- 18. In this design, they are housed in the top of the device housing 312 that is mounted to the XYZ gantry 48. Each Arduino 340 monitors and controls 6 detection channels of the apparatus 300, so that there is one Arduino for each row 342 of detection cells 302. The 4 Arduinos 340 are mounted inside 344 the housing 312. Each Arduino 340 powers a single UV LED light source 321 that is split into 6 different equal beams. Each of these 6 beams is then used to perform the absorbance measurement of each of 6 fluid samples for a selected row 342 of detection cells.

FIG. 18 shows at top view of the housing with the 4 Arduinos slotted in place. The UV LED is connected with 3 leads 323 and rests in place where it is connected to the fiber optic cable that is split into 6 equal beams as described above.

The apparatus 300 is controlled by software which provides a user interface to allow the user to control its various functions. The software may be stand alone software i.e., the software provided with the controllers 40. Preferably however, the software for operating the apparatus 300 is integrated with software 30 so that monitoring absorbance of the various samples may be integrated with other functions of the protein purification device. A user 101 may communicate with software 30 such as Crystal Monitor through a graphical user interface (GUI) driver 304, the driver 304 generating additional screen shots to allow for monitoring and controlling the apparatus 300 as discussed below.

Given the complexity of simultaneously measuring and monitoring 24 UV channels, the apparatus 300 of the invention has a user interface 304 that is simple and intuitive to use. Below is a brief description of the software along with screen shots of the actual interface 304.

FIG. 19 is a screen shot of the main user interface page as discussed above. The different methods that users 101 have developed are listed and can be searched by name, author, date, etc., allowing for duplication of a particular trial. FIG. 20 is a screen shot of the deck configuration page. It is here that the user 101 sets positions for fraction collection and defines X, Y, and Z coordinates for the gantry' s 48 movement.

FIG. 21 is the Column Channel Group tab. This tab enables the user 101 to group different channels together for performing specific actions. With 24 different channels and 24 samples to keep track of, simple and customizable grouping options are critical to the ease of use of the user interface.

FIG. 22 is a screen shot of the Manual Control tab. Here the user 101 can execute single and specific movements of the gantry and pumps. This page also includes a live readout (top two rows) from each of the 24 detection cells 302 so the user 101 can monitor the measurements coming out from each the channels.

The UV measurements are collected and organized by the User Interface 103 via GUI 304 and displayed in a manner common and recognizable to protein scientists. FIG. 23 is a screen shot of the real time UV readout tab. There is also an overview mode so that the user can see the latest activity.

From the foregoing description, one skilled in the art can easily ascertain the essential characteristics of this invention and, without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

It is to be understood that the present invention is not limited to the sole embodiment described above, but encompasses any and all embodiments within the scope of the following claims:

Claims

CLAIMS I claim:

1. A system for monitoring, mixing, and delivering solutions, comprising: a plurality of precision pumps, each pump drawing an associated stock solution from a solution source, and pumping the drawn solution out through an outlet via a conduit, each of said conduits fluidly coupled to one of said precision pumps; a distributor which directs a solution from a particular conduit to a selected one of a plurality of solution holders; a UV monitoring component having a plurality of UV detectors for monitoring absorbance of fluid in said conduits; and, processor systems for controlling the pumps, the distributor, and the UV detectors.

2. The system of claim 1, further comprising: a multi-port distribution valve associated with each precision pump, each valve for connecting its associated pump to one of a plurality of associated inlets and outlets, the processor system controlling each valve.

3. The system of claim 2, individual inlets of a particular pump being connected to different solutions, and each outlet of said pump being uniquely associated with one of said inlets, such that a particular solution always enters through one of said inlets and always exits through the associated outlet.

4. An apparatus for simultaneously monitoring an array of fluid handling conduits comprising:

An array of fluid receiving detection cells in fluid communication with said array of fluid handling conduits, each of said detection cells having a flow cell with a sensor positioned therein, said flow cell allowing for transmission of electromagnetic energy therethrough; a source of electromagnetic energy coupled to an array of transmission lines, each of said transmission lines terminating at one of said detection cells such that the output of said transmission line radiates through said flow cell and onto said sensor; and, a processor for receiving an electrical output from each of said detection cells, a display associated with said processor for displaying data related to said detection cell output.