CA2192582A1 - Device for packing chromatographic stationary phases - Google Patents

Device for packing chromatographic stationary phases

Info

Publication number
CA2192582A1
CA2192582A1 CA002192582A CA2192582A CA2192582A1 CA 2192582 A1 CA2192582 A1 CA 2192582A1 CA 002192582 A CA002192582 A CA 002192582A CA 2192582 A CA2192582 A CA 2192582A CA 2192582 A1 CA2192582 A1 CA 2192582A1
Authority
CA
Canada
Prior art keywords
column
stationary phase
continuous
passageway
funnel
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
CA002192582A
Other languages
French (fr)
Inventor
Michael R. Ladisch
Kent Hamaker
Richard Hendrickson
Mark Brewer
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Purdue Research Foundation
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of CA2192582A1 publication Critical patent/CA2192582A1/en
Abandoned legal-status Critical Current

Links

Classifications

    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01DSEPARATION
    • B01D15/00Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08Selective adsorption, e.g. chromatography
    • B01D15/10Selective adsorption, e.g. chromatography characterised by constructional or operational features
    • B01D15/20Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the sorbent material
    • B01D15/206Packing or coating
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/50Conditioning of the sorbent material or stationary liquid
    • G01N30/56Packing methods or coating methods
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/50Conditioning of the sorbent material or stationary liquid
    • G01N30/56Packing methods or coating methods
    • G01N2030/562Packing methods or coating methods packing

Abstract

Devices (10) and methods for packing a chromatography column with a continuous stationary phase. The preferred device (10) includes a brace member (11) with an engagement surface (12). The brace member (11) defines a thru-hole (13) at a location so that a passageway defined through the column is aligned with the thru-hole (13) of the brace member (11) when a first end of the column is engaged with the engagement surface (12) of the brace member (11). A funnel member engageable with an end of the column opposite the first end. The funnel member has converging walls configured to radially compress the continuous stationary phase for entry into the passageway of the column when the continuous stationary phase is pulled through the funnel member. The device (10) includes a mechanism for applying mechanical force (45) to pull the rolled continuous stationary phase through the funnel member wherein the stationary phase is radially compressed and enters the passageway of the column.

Description

WO~13~361 PCTI~IS9~107018 ~192582 DEVICE FOR P~CK]NG CHROMATOGRAPHIC
STA'l'IONARY Pi~.SES

This inverltion was made witll Government support under Grant No. BCS ~912150, awarded by the National Science Foundation. The governrnent has certain rights in the inverltion .

FI~LD C)F I'IIE INVrN'i'ION

rhe preserlt inverltion broarll~ concerns chromatograE~ily, paLticularly, stationary phases for use in chrornatography.
MoLe speciLically, tlle invellLion concerns devices and m~thods for packillg cllromatograpily columns with continuous stationary E~hases.

BACKGROUND OF TIIE INVEll'rION

Chromatograpl-y is one of the rnost widely used analytical techniques in biotechnolog~. Currently, separation costs account for well over 40% of the production e~cpenses associated with the manufacture of proteins and other bio~ecllnology products. Key objectives towards managirlg these costs include reducing tile number or comple~ity of production s~eps and labor costs.

Parliculate supports are ~he most commorlly used materials for chromatograpilic sLationar~ phases. ilowever, these materials are inefficient for large scale operatiorls which require speed, scale up and ease of clean operation. On the otller han~, contirluous stationary phases formed of fabric !

~VO95/3~361 PCT!1!595107018 2~925~2 (i.e~ a textile structure composed of mechanically interlocked fibers or filaments) provide many advantages compared to discrete spsherical or granular packing materials, including: mechanical stability at extremely hi~311 eluent S linear velocities, wide availability of a range of definecl surface chesrlistries in large quantities, and constant plate height at high linear velocities, thereby making scale-up and operation over a wide range of conditions relatively independes1t of flow rate. Furt1lerrnore, the fabric, when packeo tigsltly in column, retains the advantage of low pressure ùrops, which is dif~icult to achieve wit parLiculate forms of the same materials.

Continuous stationary phases have further advantages ove.r loose packing materials, including, very fast flow rates, }5 direct scale-up, and clean columns. These advantages are not currently found in preparative and commercial scale chromatograehy using particulate cellulosic stationary phases.

In spite of these advantages, columns having contin1lous pslases of fabric are difficult to pac~ with sufficient densitiT and reproducibility. A need thus exists for devices and methods which provide reproducible, dense packing of a large number of columns at reasonable e~pense. The present invention addresses tslis need.

WO9513~361 PCT111S95/07Ulg 21~2~82 SUM~ARY OF TIIE INVENTION

In accordance with the inverltion, devices and methods are provided for packing a chromatograpl1y column with a continuous stationary phase. Briefly mentioning one aspect of the invention, such a device includes a brace member having an engagement surface for engaging a column to be packed. The brace member also defines a hole extendiny completely through the brace member ~hereinafter "thru-hole"~
at a location so that the passagewa~ defined through the colurnn is aligned with the thru-hole of the brace member when an end of the column is ensased with the engagement surface.
T11e device also includes a ~unnel member engageable with an end of the column opposite the engaged end, wherein the funnel member has converging walls configured to radially compress the continuous stationar~ phase for entry into the passac1eway of the column when the continuous stationary phase is pulled through the funnel member. A mechanism is also provided for applying mechanical force to pu11 the rolled contir1uous stationary phase through the funnel member wherein the stationary phase is radially compressed upon entering the passageway of the column.

Anoti1er preferred entbodimer1t of the invention provides a process for packing a chromatography column with a continuous stationary phase. The inventive process comprises the step 2s of providing a continuous stationary phase and a column having a passageway therethrougll, wherein the passageway is sized to receive the continuous stationary phase upon radial compression o~ the continuous stationary phase. A funnel member is provided having converging walls configured to radially compress the COIltil1UOus stationary phase upon passage of the stationary phase through the funr1el member, In accordance with the invention, the continuous stationary phase is positioned within the passageway of the column with W0~5/3~36] rcr~ls~s/l~70l8 ~192~7~ 4_ applied mechanical force, wnerein the positionilly includes passage of the continuous stationaLy phase through the funrlel member so as to radially compress the continuous stationary phase, as it is being pulled or packed into the colurrn, One object of t.he invelltion is to provide a device which reliably and efficiently packs continuous stationar}7 phase rnaterial into c~lro~atograpi-y colurnns.

Anotiler object of the present iuvention is to provide methods for yuickly and reproducibly packing continuous Iq .~tationary phase material into chromatography colurllrls.

Additional objects, features and advantages o~ the invention will be apparent from tne following description.

WO9S/343~1 2 1 9 2 5 ~ 2 PCT~S9510701X

BRIEE' DESCRIPTION OF THE DRA~IINGS

Figure l is an exploded perspective view of a device of the present invention.

Figure 2 is a side cross-sectiorlal view of a device connected to a rolled, continuous stationary p11ase.

Figure 3 is a side cross-sectional view of the funnel member used for column packing.

Figure 4 is a side cross-sectional view of .3 brace member, stopper, and column.

Figures 5 (A)-(C) show tlle progression of rolled stationary phase into a column according to the invention.

Figures 6 (A)-(C) show side cross-sectiorlal views of the colwnn, stopper, and funnel assembly during a packing sequence.

~r'O 9S13;i361 . PCv1'111~:}'3~.~071ll#
2~$2~82 ~ESCRlPTIO~ OF l'llE PREFERRED EMBODlMENT

F'or the purposes of promoting an understanding of trhe pr~ ciyles of the invention, reference will now be made to tlle preferred emb~odilllents thereof, and specific language will be used to describe the same. It ~rill nevertheless be understood that no limitation of the scope of the invention is ~heIeby intended, such alterations, modifications, and further applications of the principles of the inventiorl being contemplated as would normally OCCUL to one skilled in the l(l art to which trhe invention relates.

l'he present invention ~rovides devices and methods for ;-ackiny a chromatograpl-y column, such as a liquid or gas chronlotagrapily column, wiiil a continuous stationary phase.
~'he invention thus enables the exploitation of the advantages of continuous stationary phase materials in large scale operations. Corntirluous stationary phase materials are excellent sorhents and can be effectively derivati~.ed to improve tlleir soIbent properties. For example, contilluous statiorlary phases foImed wiLh materials Euch as cotton and otller cellulosic niaterials have proven highly effective in the separation oE pIoteills and other substances.

One preferred embodiment of the present invention provides a device 10 for packing chromatography colurr~ns StlCn as that depicted in FIGS. 1 and 2. The preferred device 10 2s includes a brace niember 11 having an engagement surface 12 for engaging or supporting an end of a chromatography column 20, a funnel member 33 having converging waLls 34 (~IG. 3) at a flare angle configured to radially compress the continuous stationaLy phase of ifabric 60 For entry into the colurmrl 20, anc'i a mechanism for applyirlc3 mechallical Lorce to pull the fabric ~o tllrougil the ifuunel and into the columrl 20. As usecl herein, the term flare angle refers to the angle defined ~095/3U6l ~1 9 2 5 8 2 PCT/liS95/0701 between the converging walls 34 and the lonyitudinal axis "A"
(Fig. 3) of the funnel member. The brace member 11 and the means for applying mechanical force are configured to facilitate pulling the phase 60 into the column 20.

The brace member 11 defines a thru-hole 13 at a location such that a passageway 23 defined through the column 20 can be aligned with the thru-hole 13 of the brace member 11 when the end 21 of columll 20 is enga9ed with the engagement surface 12 of the brace member 11. I'he brace member 11 is preferably composed of a material such as metal or plastic, for exarnple steel, whicll is strong enough to resist movement or mecharlical failure when subjected to the forces necessary to pull the stationary phase into the column 20.

The preferred device also includes a funnel member 33 (~IG. 3) which is engageable with the end 22 of the column 20 which is opposite the end 21 engaged with engagement surface 12. As mentioned previously, the preferred funnel member 33 has converging walls 34, which generally form a conical section and whicil provide a flare angle configured to radially compress the fabric 60 for entry into the passageway of the column 20 when the fabric 60 is pulled through the funnel member 33 in the direction of convergence of walls 34. E'referably, the colunm 20 is cylindrical and the diameter of the continuous stationary phase 60 is reduced to Zs a diameter slightly less than the inner diameter (I.D.) of the column 20 as it is radially compressed. The flare angle employed will depend upon the bulk density and other properties, such as tensile strength, of the continuous stationary pllase 60, as well as the packing density desired, and can be chosen to facilitate a smootll transition of the phase 60 into the column 20.

~~'0 g~/3S3~1 PC~TII,IS95,'07(~1~

2192~8~ -8-'rhe column-engaging end 36 of funnel member 33 preferably has an I.D. slightly greater ~han the outer diameter (O.L').) of the colurnn 20, so that the column 20 is receivable in the engaging end 36 of the funnel mernber 33. In ~ is manner, and S with the converging walls 34 converging to a diameter essentially equal to the I.D. of column 20, a smooth surface transitioll between the walls 34 and the interior walls of the column 20 is provided to facilitate passage of the contilluou3 stationary phase 60 into the column 20 wL-ile avoiding damage ~O ~L~e pLIase 60.

In the preferred device 10, tlle brace member 11 is held in spaced relationship relative to a mechanism for applyiny rnechanical force to pull trhe rolled continuous stationary pllase 60 throLIgh the funnel member 33 and into the column lS 20. I'he inverltion contemplates employment of any one of several means for applyiny mechanical force wllicll are well knowll in the art. ~or e~ample, mecilanical force can be applied via manual operation of a mechanism whicrl leverages tLIe applied manual force so as to increase the same and direct the increased force to the positioning of the stationary phase 60 into the column 20. In the illustratecl embodilnerlt, a hancl drawn wincll 45 is utilized, although other means such as levers, screw drives or the like can also be used. It is also contemplated that an electric or hytlraulically driven motor or winch could also be employed tc, cleliver mecllarlicrll force instea~ of the halld oeerated wincL
45 or other manually-operated means.

Referring to FIGs. 1 arld 2, a llalld drawn wincl3 45 and a cord member 44 are operable to pull the rolled continuous stationary phase i,0 through the funnel member 33 and into the passac3eway of coluinll 20. In one embodiment the winch 45 and brace member 11 (optionally with appropriate spacers such as W095l3~36l PCTf~S9SfO7018 2192~82 _9_ llA) are bolted to a base 16 by mountiny bolts 17. I'he cord memiJer 44 preferably includes a cable 47 havilly a first end engaged to the winch 45 and a second end 49 engaged to wire 50 which is threadable through the thru-hole 13 of the brace member, the passageway of a column 20, and the funnel member 33. The wire 50 is attachable to tne continuous stationary phase 60. A connecting device, such as a hook 48, is attached to Lhe second end 4g of the cable 47 for connectirly to the wire 50. 1'he cable 47 can be constructed from any suitable material having higll tensile strength and resisting elongation under the ~orces aE)plied~ for example a metal such as stainless steel, or a polymeric material such as Kevlar~
aromatic polyamide.

According to a preferred embodiment, the device 10 for lS packing continuous stationary phases also includes a stopper member 25 having a bore 30 therethrough. The stopper member 25 aids in aligning the passageway 23 of the column 20 with the thru-hole 13 in the brace member 11 and therefore aligns ~he continuous stationary pllase 60 with the directional pull of the winch 45. Such alignment minimizes shearillg of or other damage to the continuous stationary phase 60 as it is positioned into the column 20.

Referring now to FIG. 4, the stopper member 25 preferab]y has a first bore 27 having an I.D. equivalent to the I.D. of 2s column 20 and a second bore 28 having an I.D. slightly greater than the O.D. of column 20 so that column 20 is receivable in the second bore 28 of the stopper member 25.
1'his permits entry of the column 20 into the stopper menlber 25 to sbut interior walls defined at the transition between tlle first and second bores 27 and 28, as illustrated. ln the preferred embodirnent, the stopper member 25 is a separate Ullit. In this manner, the packing of columns of differing outside diameters can be facilitated by selecting and ~09513~361 ~ PCT~S~ 7018 2192~

employing a stopper member 25 havin~J the appropriate dimensions.

In the preferred device, tlle first end 29 of the stopper member 25 contacts tile engagement surface lZ of the orace member ]1 at a location such that the passageway 23 of colum 20 is aliqned and in con~nullicatiorl with the thru-llole 13 of the brace member 11 when an end of column 20 i.9 inse.rted in the bore 30 of the stopper member 25. In this arrangement employing the stopper lllember 25, column 20 is of course ultimately engaged to the engagemerlt surface via the stoppe rnerrlber 25.

I'he engagemerlt surface 12 of the brace member 11 is preferably adapted to secure the stopper member 25 in place wherl the two components are engaged with one another. For exarrple, flanges or similar protnrbances can be pro~;ded in a spaced relationship around the thru-tlole 13 of a brace member 11, and configured to secure tlle stopper member 25 in position against the engagerllent surface lZ. Preferably, thouyh, a recess is defined in the engagement surface lZ
about the thru-i~ole 13, the recess being climensioned to receive the end of the stopper member 25 as illustrated (FI~.
4).

The stopper member 25 and funnel member 33 can be machined or otherwise formed from suitable metal (e.g.
~5 stainless steel or alumiuum) stock or from standard chromatograplly columrl enrl-fittings which correspond to tile V.~. of a column of interest, for example 3f5", 1~2", etc.
E~or example, such fittings are commercially available under lhe bralld nallle ~alco~.

:JO The colunm 20 may be composed of any suitable material known in the art such as glass or stainless steel (e.g. 316) WO95/3J36l PCT~95/0701 n~ro~
JO~

seamless tubirlg. Because of the extrerne tensile force and friction between the column 20 walls and the stationary phase 60 during packing, it is pre~erable to remove snags, burrs, or other anomolies preserlt inside or at the ends of the columns 20 prior to the packing procedure. The colurnns 20 can be polished at both ends using a hiyll speed lathe.
Columns 20 are also preferably conditioned against corrosion by pacifying the inner surfaces by Lreatment with an acidic solution such as concentrated nitric acid, for example for a period of 30 minutes, followed by rinsing with water.
Additionally, prior to packing, the interior of columns 20 are preLerably wiLIl an appro~riate lubricant, for example polyethylene glycol, M.W. 200 (PEG 200) to reduce friction durinq the packing operation.

To condition the stationary phase 60 for aLtachment to wire 50, it is preferably pierced at one end with, for example, an awl or a {elatively small, e.g. 1~16", high speed drill. The pierced hole 63 is then threaded with a length of wire 50. The wire 50 preferably has a small diameter and high tensile strength. The preferred material is Kevlar~
aromatic polyamide fiber, which is non-abrasive and has a greater resistance of elongation than steel. A loop or loops of wire 50 are then threaded through the funnel, column 20, stopper member 25 and thell the thru-!!ole 13 of brace member 11. The loop or loops are therl secured to the hook 48 or other connecting device.

Tlle contirluous stationary phase 60 is preLerably lubricated before being packing into the column 20. The lubricant can be cllosen Lrorn lubricants well knowll in the art, for example, polyethyleneglycol such as PE~ 200.

Referring to FIGS. 5 and 6, shown are drawings whicll ~urther illustrate the packing operation. An engaging end 36 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ , . ... ..... .. _ . .. _ . . . _ .... _ WO95~3~361 ~Pcr~s~m7ulx 21~25~ ~

oL the funnel memher 33 is engaged to the column 20 and the column 20 is engaged to tlle engagemerlt surface 12 of the brace member 11. As previously de5cribed in the preferred operation the column 20 is engaged to ts~e surface 12 via s stopper melllber 25. The wire 50 threaded throuqh the otller components and attached to the stationary pnase 60 as discussed above is also connected to the hook 48. In tu{n ~he hook 48 is colmected to tlle cable ~7 and thus ultirnately to the wincll 45. Rotating the handle 46 to oQerate the winch 45 pulls Ihe stationary phase 60 tllrourJh funnel member 33 and into and throllgll the column 20. This packing operatioll is preferably executed b~ applying a slow steady force on the pha.se 60 as it proceeds throuyll the funnel member 33 and tne columrl 20. As well it is preferred to slightly t~sist the stationary phase 60 in the direction it was rolled to facilitate ttle packing proce~ure. In addition it is preferred that the stationary phase 60 be secured for hatldling using plastic ties 62 or other securing means (such as Velcro~ hook and loop fastener strips) positioned along ~U the entire length o~ the pha~e 60. The ties 62 can be removed e.g. cut off as they near the column 20 as the pl~ase 60 is pulled into anrl throuc~tl the column 20.

The stationary phase 60 is preferabl~ pulled througlL the colusnrl 20 so that a portion of the phase 60 can he observed protruding througll Lhe through hole 13. It is desirable that tlle poLtiorl of stationary phase 60 containing the hole 63 clear the end of the colurml 20 so that it can be removed.
Once the rolled fabrsc 60 is pulled through the column 20 the device 10 is relieved of Lension and the stopper mernbeL
3U 25 and funnel member 33 can be removed. The column can then be finished by cutting the corltinuous stationary phase 60 flush with the end of the colurnn 20 using a sharp insit-urnent SUCII as a razor blade. ~n appropriate stansdard liquid chrrJmatograplly columll end-fittillg can then be fitted to l:he WO9513~361 PCTA!595/07018 ~ 2192~8~

colum11 20. Any lubLical1t utili~ed in the packing operation ~ can be washed from the stationary phase with an appropriate eluent after completion of the packing operation.

If the rolled stationary phase 60 must be later removed from the column 20, a punch with an O.D. correspondin~ to the I.i~. of a column 20 can be used. Alternatively, the packed column can be pumped full of lubricant such as PEG 200, and the stationary phase pushed or pumped from the column. This enables the valuable statio1lary phase 60 to be retrieved wit minimal damage.

In a preferred aspect oE tl1e inver1tior1, the contir1uous stationary phase 60 can be a slleet of continuous stationary pi1ase material rolled into a cylindri.cal configuration. It is contemplated that this rollin~ step may be manual or automated. Materials for the continuous stationary phase can be chosell from various types of materials which are within tlle an1bit of the skilled artisan. Preferably, the stationary pt1ase is formed from a rectangular or square sheet of fabric. i~ectangular fabric can be oriented in a ~arp parallel, fill parallel or bias direction. The length of the fai~ric in tt1e direction of flow within the colum1l will be selected depending on the desired lengt11 of the column. The widtl1 and thickness of the fabric will, at least in part, determir1e the diameter of the roll of fabric and hence the extent of radial compression necessary to position the material in a given column diameter. Preferred stationary pt1ases are prepared as described in a copending U.S. Patent Application of Michael Ladisch et al. entitled NOVEL METIIOD
FOR DERIVATIZATION OF CELLULOSE STATIOi~ARY PHASES, filed June l5, 1994, whic1l is hereby incorporated herei11 by refeLence in its entirety.

~Voss/3~36l ' PC~'/~1~9~)7018 2i925~ ~

l'he stationary phase is preferably rolled to a cylinrlrical configuration on a flat, hard and smooth sùrface, be9i.11nillg Wit}l the smallest uniform fold possible. The fabric is rolled by hand, or by a mechanical device so that Lhe entire lengtll oE the Eabric, the lengLh to be perpendicular to flow, is in a cylindrical configuratioll.
The tightness of the roll can be increased by repeatedly ro]ling the stationary phase across a table or other suitable surface using the palms of the hands, wllile exertirlg an increasir!gly stronger dowllward force on the roll. C~nce sui~icient tightness or com~action of the roll has been achieved, the roll can be secured for handling using plastic ties 62 or other securing mealls as described above.

Preferred colllmrls prepared in accordance with the invelltion, contairting rolled cotton or a similar rolled cellulosic fabric, can have adYarltageous void Lractions and packing densities. For example, advantageously high packing densities of greater than about 0.5 grams/cc can be achieved in the column, typically falling within the range of about Z0 0.5 g/cc to about 0.6 g/cc. Advanta~eously low void fractions of about 0.~ or less, or even about 0.~ or less (as rneasured by bovine serurn albumin, BSA), can also be readily achieved in accordance with the invention. Columns of the in~Tention exhibit advantageousl~ loh pressure drops and stability against mechanical breakdown as demonstrated by the observance of consistent pressure drops over extended periods of use. As well, columns prepared in accordance with the inverltiorlllave advantageous flow properties, for example readil;7 providing linear flow velocities in excess of 50 ~0 cm~min and ranging up to about 100 cm/min.

Moreover, using rolled fabric stationary phases in the inverltion provides the bene~icial ability to pach large quarltities of fabric into co'iumns in a manner giving the higl ~09513~361 21 9 2 ~ 8 ~ PCT/IT~95/07018 density desired while avoiding more laboriou.s met11ods such a.s tl1ose which would require the cuttiny, stacking and pac~.inc3 of discs of fabric. Moreover, the use of stacked discs and - attendent cutting operations will invariably lead to significant amounts o~ wasLe stationar~ EJhase and thus overall increases in procluction costs. As well, physical lin1itations in effectively and reproduceably stacking discs, such as those leading to limitations in colu1ml heicJllt, may be avoided in accordance with the invention.

While tne invention has been illustrated and describeù in detail in the drawinys ancl foregoing description, the same is to be considered as illustrative and not restrictive in character, it beiny uudersLood tllat only t}le preferred embodime11t has been shown and descri.bed and that all chanyes lS and modifications that come wichin the spirit of t11e iuventio1l are desired to be protected.

Claims (32)

What is claimed is:
1. A device for packing a liquid chromatography column with a continuous stationary phase, comprising:
a brace member having all engagement surface, the brace member defining a thru-hole at a location so that a passageway defined through the column is aligned with the thru-hole of the brace member when a first end of the column is engaged with the engagement surface of the brace member;
a funnel member engageable with an end of the column opposite the first end, the funnel member having converging walls configured to radially compress the continuous stationary phase for entry into the passageway of the column when the continuous stationary phase is pulled through the funnel member;
a mechanism for pulling the stationary phase through the funnel member;
a cord member attached to the mechanism, the cord member being threadable through the thru-hole, the passageway and the funnel member and attachable to the continuous stationary phase; and the mechanism and the cord member operable to pull the rolled continuous stationary phase through the funnel member wherein the stationary phase is radially compressed, and into the passageway of the column.
2. The device of claim 1 wherein:
the mechanism comprises a winch held in spaced relationship to the brace member
3. The device of claim 2 wherein the cord member comprises aromatic polyamide fiber.
4. The device of claim 1 further comprising a stopper member defining a bore therethrough, wherein the column is receivable is the bore of the stopper member, and the stopper member is engagable with the engagement surface of the brace member at a location so that the passageway in the column is in communication with the thru-hole of the brace member when an end of the column is received in the bore of the stopper.
5. The device of claim 9 wherein the engagement surface of the brace member comprises flanges spaced around the thru-hole of the brace member, the flanges being configured to secure the location of the stopper member when engaged with the engagement surface.
6. The device of claim 4 wherein the engagement surface includes a recess about the thru-hole, the recess dimensioned to receive and fix the location of the stopper when engaged with the engagement surface.
7. The device of claim 4 wherein:
the thru-hole of the brace member is aligned with a directional pull of the winch; and the stopper member is configured to align a longitudinal axis of the column with the thru-hole of the brace and thus also with the directional pull of the winch.
8. The device of claim 1 wherein the funnel member is engageable to the column at an engaging end of the funnel member the engaging end being dimensioned to receive an end of the column.
9. The device of claim 1 wherein the engaging end of the funnel member is cylindrical in shape and has an inner diameter sized to receive a cyclindrical column.
10. A process for packing a chromatography column with a continuous stationary phase, comprising:
providing a continuous stationary phase;
providing a column having a passageway therethrough, the passageway being sized to receive the continuous stationary phase upon radial compression of the continuous stationary phase;
providing a funnel member having converging walls configured to radially compress the continuous stationary phase upon passage of the stationary phase through the funnel member;
positioning the continuous stationary phase within the passageway of the column with applied mechanical force, said positioning including passage of the continuous stationary phase through the funnel member so as to radially compress the continuous stationary phase.
11. The method of claim 10 wherein the continuous stationary phase is cylindrical in shape.
12. The method of claim 11 wherein the continuous stationary phase is a rolled fabric.
13. The method of claim 10 wherein said positioning includes pulling the continuous stationary phase into the passageway of the column.
14. The method of claim 13 wherein said positioning includes:
attaching a cord member to the continuous stationary phase;
threading the cord member through the funnel member and the passageway of the column;
engaging the cord member to a winch; and operating the winch to pull the continuous stationary phase through the funnel member and into the passageway.
15. The method of claim 14 wherein said attaching includes threading the cord member through a hole in the continuous stationary phase.
16. The method of claim 10 further comprising lubricating the continuous stationary phase before positioning the continuous stationary phase within the column.
17. A process for packing a chromatography column with a continuous stationary phase, comprising:
(i) providing a device including, a brace member having an engagement surface, the brace member defining a thru-hole at a location so that a passageway defined through the column is aligned with the thru-hole of the brace member when a first end of the column is engaged with the engagement surface of the brace member, a funnel member engageable with an end of the column opposite the first end, the funnel member having converging walls configured to radially compress the continuous stationary phase for entry into the passageway of the column when the continuous stationary phase is pulled through the funnel member, a mechanism for applying mechanical force to pull the rolled continuous stationary phase through the funnel member wherein the stationary phase is radially compressed, and into the passageway of the column;
(ii) engaging the first end of the column to the engaging surface;
(iii) engaging a first end of the funnel member to the end of the column opposite the first end of the column;
(iv) positioning a continuous stationary phase at a second end of the funnel member opposite the first end of the funnel member; and (v) operating said mechanism to apply mechanical force to the continuous stationary phase to pull the stationary phase through the funnel, wherein the stationary phase is radially compressed, and into the column.
18. The process of claim 17 wherein:
said mechanism for applying mechanical force includes a winch and a cord member attached to the winch and the stationary phase.
19. The process of claim 18 wherein the cord member is threaded through a hole in the continuous stationary phase.
20. The process of claim 19 further comprising pulling the first end of the continuous stationary phase beyond the first end of the column, the first end of the continuous stationary phase including the hole, thereby causing the first end of the continuous stationary phase to protrude beyond the first end of the column; and removing the first end of the stationary phase from the remainder of the stationary phase.
21. The process of claim 17 further comprising lubricating the continuous stationary phase before applying the mechanical force.
22. The process of claim 18 further comprising equilibrating the continuous stationary phase in an eluent after inserting the phase in the column.
23. A chromatography column containing a continuous cellulosic stationary phase at a packing density of about 0.5 g/cc or greater.
24. The column of claim 23 wherein said packing density is in the range of about 0.5 g/cc to about 0.6 g/cc.
25. The chromatography column of claim 23 wherein said fabric comprises cotton.
26. The chromatography column of claim 25 wherein said packing density is in the range of about 0.5 g/cc to about 0.6 g/cc.
27. The chromatography column of claim 26 wherein said column has a void fraction of about 0.3 or less as measured by bovine serum albumin.
28. A chromatography column, comprising:
a column;
a stationary phase packed in said column, said stationary phase including rolled cellulosic fabric radially compressed within said column and exhibiting a packing density of at least about 0.5 g/cc;
said column further having a void fraction of less than about 0.4 as measured by bovine serum albumin.
29. The chromatography column of claim 28 which is characterized by its preparation by a process including the step of positioning the rolled cellulosic fabric within the column with applied mechanical force, wherein said positioning includes passage of the rolled cellulosic fabric through a funnel member having converging walls so as to radially compress the rolled cellulosic fabric as it enters said column.
30. The chromatography column of claim 28 wherein said column has a void fraction of less than about 0.3 as measured by bovine serum albumin.
31. The chromatography column of claim 28 wherein said fabric comprises cotton.
32. The chromatography column of claim 28, which is the product of a process including:
providing a rolled cellulosic fabric stationary phase;
providing a column having a passageway therethrough, the passageway being sized to receive the rolled cellulosic fabric stationary phase upon radial compression of the stationary phase;
providing a funnel member having converging walls configured to radially compress the rolled cellulosic fabric stationary phase upon passage of the stationary phase through the funnel member; and positioning the rolled cellulosic fabric stationary phase within the passageway of the column with applied mechanical force, said positioning including passage of the stationary phase through the funnel member so as to radially compress the stationary phase.
CA002192582A 1994-06-15 1995-06-02 Device for packing chromatographic stationary phases Abandoned CA2192582A1 (en)

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DE69524749D1 (en) 2002-01-31
EP0764043A1 (en) 1997-03-26
EP0764043B1 (en) 2001-12-19
US5770060A (en) 1998-06-23
EP0764043A4 (en) 1997-05-28
US5618434A (en) 1997-04-08
WO1995034361A1 (en) 1995-12-21

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