CA2006880A1 - Push column and chromatography method - Google Patents
Push column and chromatography methodInfo
- Publication number
- CA2006880A1 CA2006880A1 CA002006880A CA2006880A CA2006880A1 CA 2006880 A1 CA2006880 A1 CA 2006880A1 CA 002006880 A CA002006880 A CA 002006880A CA 2006880 A CA2006880 A CA 2006880A CA 2006880 A1 CA2006880 A1 CA 2006880A1
- Authority
- CA
- Canada
- Prior art keywords
- column
- sample
- set forth
- chromatography material
- device set
- 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
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/34—Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/101—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/405—Concentrating samples by adsorption or absorption
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
- G01N30/6047—Construction of the column with supporting means; Holders
Abstract
ABSTRACT
An apparatus and method for chromatography of DNA, RNA, proteins and other molecules includes the use of a column adapted to hold a chromatography material and a sample to be filtered. A
pneumatic pressure differential is applied across the column and the sample is urged through the chromatography material. A
selected portion of the sample may then be collected.
An apparatus and method for chromatography of DNA, RNA, proteins and other molecules includes the use of a column adapted to hold a chromatography material and a sample to be filtered. A
pneumatic pressure differential is applied across the column and the sample is urged through the chromatography material. A
selected portion of the sample may then be collected.
Description
G~ 3~ !6:53 ?, ~/
'; 20~6 ~a~
~85~153 Ç ;C ~ ~ Q J~ 8 /, ~ .
~ e p~e~ent lnv~n~ion relate~ l;o an spparatu~ a~d ~aethodology ~or th2 ch~o~atograp~ o~ mate~ial~, and in parti~ula~, ohro~atogr~phy b~ ed on mol~au~ar ~ , a~inlty and the like ~ used, ~o~ exampl~, in t~ purlfiQation~ ~ep~ra~iDn or i~41atior~ o~ ~NA and RNA ~ragment~, pro~Qln~ an~ other mole~ul~s.
a. ~5~
Re~oving unlnoorpo~ated nucleotide~ from DNA and ~NA
P~ag~nt6, i~ola~ing ~NA fractlons, pu~i~y~n~ protQ~ns and oth2r maoro~ oulo~, ar~ important prooedures having a v~riety o~
applicAtions. ~n ~N~ and RNA ~ynth~8i~, unlnc~r~oratQd nualoo~ld~s mu~t o~ten bQ removed wh~n ~on~trua~iny nick-tran~lat~d prob~, RNA probea and end-labeled ollgonuoleotidee, ~3 woll ~ illed-in" DNA ~ragmcnt~ i~portant ~o s~parat~ the unlncorporated ~Qe-nuelao~ides from the lab~l prob~ a~ unln~oxporated lab~l may bind to the sQlid ~uppor~, re~ulting in una~cep~ably hlg~ level~ of background nol~e.
~olat~on o~ ~NA ~actions ~a~ b~ Qmploy~d in th~ ~epara~ion of, ~
~or exa~pl~, poly~denylat~d RNA ~o~ nonpoly~danylat~d RNA~. ThQ .
u~ or ohromatoyra~hy m~thods to isolato and identi~y prot~ln~
and oth~ maoromole~ules i6 anothe~ well ~nown applicat~on. -`
Current ch~o~tog~aphy methods, u~e~ part~ularly i~
c~nnectlon w~ DNA an~ 6yn~ssls, tnclud~ ion-~xchange ahrom~tog~aphy, ~eYe~al variation~ of ~1 ch~omatography and other~. Each has i`~9 own d~advantag~s. For exa~ , ion-ex~hange ~thods ~ui~e a n~ber o~ step3 which ~y ~e~lt in a si~lfic~nt invQstment o~ ~im~ an~, in th~ ca~ o~ rad~ol~bele~
' ~ '.'' DE~, 2~'8~ i6:~^3 ~ . 5/ 6 ~9~
~s/a~s ~ ~:
nuol~otid~ ~ r~ng, ex~en~ive han~l:Ln~ o~ ~adioa~tive ~aterlal.
conve~tion~l ~el~ch~o~atogr~phy "drip~' ~olumn~ a~e ~diou~
r~uiring ~im~ to both pou~ ~n~ run~ ~p~n ~olumn~, A ~arlation o~ th4 i'drip~' column, ~r~ ~o~ewh~t ~a~3t~, but r~k r~atlo~
~xpo8~r~ and Gonta~in tion in the ~e of ra~onucl~ot~de chromatography, and m~y yie~d l~ss rQ~la~le r~sult~
A~ ~lt~natlve ~h~o~atogra~y approach whiah avo$d~ th~ ~
a~or~ment~on~d di~ficultie~ wou~d th~r~o~e ~ dQ~lrAb~e.
~8L:==~ ` `
~ h~ prQ~nt inv~ntlon i~ ~$~ct~d to an apparatu~ and ~ethod ~or ~uri~y~ng, iso~atlnq and separating ~teri~ls u~ing gel ~h~o~ato~aphy. To that end, a chroma~ography mat~rial an~ a ~mple may bs load~d into ~ column and pneu~at1q pro3~ur~ applled to uxg~ the ~ampl~ thxough ~he chromatography Pia,terial, whe~eby portion~ o~ the ~ampla may b~ aollacted by th~ chro~atog~aphy mat~rial and o~hor port~on~ exclu~d. In on~ ~bodl~ent, a posltl~e pn~u~atlc pr~sure iB prov~d~d ~nd in ~ s~cond ~mbodi~ent a n~gat~e pr~u~e i~ appl~d. Additlonally, a ~ovel ~upport etrU~tur~ ~ay be employed ~o 9upport the colu~n during ch~omatography. ~he ~ample may thus be gu~cXly and ral~ably trea~d.
F~gure 1 ~ a~ explodQd p~pective ~i~w o~ an appa~atus con~truot~d in a~co~ance wlth ~ha px~s~nt ~nv~ntlon co~prlslng ~ ~
column, prQssu~ in~uclng me~, a collectlon vial and asso~t~d ~ :
~upport ~ru~tur~
~ igura ~ 1B a ¢xo~ ct$onal view o~ ths appar~tu~ o~
F1gur~ a lo~ded poBiti ~E~, 28 8a !~
~0~i880 ~s~a~c }~efh~r~ng ~o Fi~ures 1 and 2 "~ oh~o~toq~apAy app~atus ¢onstruql:ed in accordan~e ~lth th~ E~reRe~t is)vent~or~ oomprise~ a gener~lly dl~k shaped ba~e ~0 havinc~ a palr o~ retainQr 1~ and a gen~ra~ly cyl~2~d~cal rial ho~ding as~embly 20 mo~nted th~xeon.
CQnt~ally located iTI the vial hold~y aa~e~ly 20 i~ a c~lln~ al ~ha~bar 22 ~or su~por~cing a collectlon v~l 30, ~rt~
whlch the elu~nt fro~ the oolumr~ m~y b~ coll~ted. ~he t~al 30 may bo a. dac~pped ~ppendo~f tubQ 02 otber ~ultab~ collecti~n ~an~. RQ~O~ab1Y mounted to t~ ba~ lo, and slidaa~ly engaglng ~h4 ex~erlor w~ll oS the v141 holding a~embly 20, ~g ~ gener~lly cylindrlcal c~lumn ~uppo~t as~e~bly ~0. ~he column ~upp~t a~e~bly 40 ln~lude~ a ~n~ral ~pert~rQ 42 ~or~ed ln ~he gone~ally plan~r uppar ~ux~ace 44 ~hereo~. As ~hown ~n Figure a, .;
the ~upp~t a0~em~1y ~ay hava A re~ nt aolla~ 46, ~uoh as an "0~' ~lng or th~ a, p~itloned ci~4um~rentially ad~ac~t the apexture 42, and a oollar re~alner 48 adapted t~ retain the coll~r ~¢ ad~a¢~nt ~he aper~ure 420 Al~ernatlYely, ~ ~hown in F1gU~G 1, th~ ~ollar 46 and the retainer 48 ~y bs ell~i~ated, optionally, a gene~ally a~lindrlcal pre6~ur~ induciny mean~
support as~e~ly 50 may be re~ovably mounted on the base 10. ~e ~.-support as~embly ma~ aompr~se a ¢~ntral a~rturs S2 ~on~ed 1~ the ~-~
g~ne~ally plan~r ~pper ~ur~¢~ B4 ther~o~, and 1~ ~on~igure~ to 61idsably engage tha ~xterio~ w~ll o~ the ~olu~ ~upport ~s~e~bly 40. ~he ~p~rt~e S~ i~ pr~rably ax~lly al~gned with th~ :~
apertuxe 4~ in the colu~n suppost sk~ucture 40, w~tch ~tself i6 .:
pre~e~s~ly axially sligned wi~h the ch~ber 22 in th~ vial holdin~ a~8~b1y 20.
~", 28 8~ ^ 5 . P. 1/14 ~006~
P~
18~7.
Al~er~ati~ , a~ ~hown $n Fi~U~çl 1., the~ 3uppor~ a~6am~1y 50 mBy ~nclude ~n upp~ u~Sac~ 54 ha~ing ro aperture t~erein. The ~upport ~embly 50 ma~ be ~ur~hex pro~ided with a pai~e o~
loa)ciJ~g tab~ 5~ adap~e~ to engage the ro~21in~rs 12 on th~ ~a~e 10 to lnotc th~ suppozt a~oe~nbly 50 ih p~a~:a du~in~ U8~. Oth~
sui'c~ble locXing me~hani~ms, such ~ thre~ds, could al~o be e~apl~yed. The assemblie~ ~0, 4~ ~nd 50 ~ay l:~e ~ormod o~ a radlatlon shlelding ~ate~l~l or, pre~erab~y, are ~on~t~uot~d to ~lt ~curely inside a bet~ ~hi~ld d~ ~loa. Molded P1A8tia nlateri~l~ have ~Q~n ~ound ~ult~ble although o~r mat~rial~ may ~1BO ~e ~snploy~d .
Supported ~y tlle column holding as~e~bly 40 ~bove l~he ~lal 30 i~ a ~ub~tsntially ~ubul~ oh~omatog~aphy column ~0. ~h~
colu~n 60 may b~ about 1 ~1 $n sizé, h~ing ~ p~a~erred lnta~nal d~amet~r o~ abo~t 5~ and a p~e~e~r~d length o~ a~out lOO~m, anq ~o~pri~e~ open~ngo ~a and 64, r~3p~ct$~ely, at each ~nd the~o~
An annula~ lip ~6 ~y b~ prov$ded olrcum~eren~ially ~d~acent ~h~
upp~r op~ning 62, a~ 5hown in Flgu~a 1. The upp~r ope~ln~ 62 is adapt~d to re~ei~o ~ ~romatog~p~y ~aterlal 70 and a 6~ple 80 to be flltered. ~he low~r op~ning 54 ~s ~ a~sa of rsduGed cro~ ection ad~pted to pre~ent pa~sag~ of th~ chro~ato~ap~y mat~r~al 70 w~le pa~t~$ng p~ a of the ~amplQ 80. ~`-Add~tlonally, a ecr~n or ~ er ~8, co~pr~sin~, ~or example, gla~ wool, may be employ~ to ~ntaln the Ghro~atogra~hY ~at~ial :~
70 withln ~he colu~n ~o. Pre~ra~ o~ ~ tubular ~olu~n, the -int~rnal dia~t~ ~hould not exceed abou~ lo~ ~or ~o~
~hromoto~r~phy appll~a~on6, lest the su~oe t~ion o~ the sample ~ in~u~ficlent to ~r~v~nt ~farve~o~nce ~nd cons~guent lo~ o~ p~e~s~Q t~rou~h tho samplo. In~reasing ~he len~ og ~ha colu~n 60 ~hould ~nhanc~ ~h~ deg~ae o~ ~par~tlon.
DE~, 28 8g liv:~t , P. 2~ l ' , . t_, j ~00688 PA~
~8S~t Pr~e~ably, the co1umr~ should b~ ns 14~ ~han ~bout 60mm in 1ength.
Pneuma~ic pre~sure ~nducing me~lh~ 94, ill thi~ ~a~e a syriTIge~ raay ~IQ at~ached to the ~lpp~r oper~ng ~2 o~ the Gol-~mn 60. A~ ~hown ~n ~igu~ 1, th~ sy~in~e ~0 may include a ~e~1e~ oS
'ch~ad~ ~2 w~1ch engage the annu1aP lip 66 o~ ~h~ colu~ 60 to . ~etaln the colu~n ~0 and th~ ~y~ln~a 90 in mutu41 engay~ment. In ~ ~1r3t ~m~od~ment o~ the inven~c10n, the syr:Ln~ gO, ~v~ng t~e plung~r 94 wlthdrawn as ~hown ~2~ F~g-lrs 2, m~y be a~taohed ~o ~he ~olu20n 60 an~ a po~tive pnsu~atio press~re d1~ferential ~ppl~ed between the open~ng~ 62 a~d 64 o~ the ~olu~n ~0 ~y d~pree~in~ t~
plun~er. In tha~ cas~, the support a0~embly 50 ~ay b~ place~
o~er the ~u~port as~ly 40. A~ the asse~ly 50 ls lowe~ B
upper ~ur~ce 5~ nc~ ap~rtur~ 52 i~ provl~ed) ~ill contaa~ ~ha plung~r 94 and auto~at~ally d~re~s ~am~ until the bott~ o~ the ~3~ppo~ 4em~1y SO Jneets thu base 10. ~t ~}~at polnt the 3upport 21~8~1~11bly 50 ~ay be ~iated unt$1 the locking tab~ 54 ~t~ t~rith ~;
the r~ta~n~r~ 1~. r~ an ~p~rture 52 ~L~ pro~id~d 1~ tbo ~upport ~esembly 50, t~ pl~ng~ 94 wlll ~x'eend there~hrough and may ~
~anually depre~d. ~n an alt~rnati~re embod~ont, not shown, the prQ~sure indu~lng mean~ 90 may be attached to the lower open~n~
~4 o~ thq colu~n ~th~ ~olu~n may be re~nove~ fro~ the ~upE:or~ -~se~bly 40 ln ~hat ~:a~e) and t~e sampls drawrl throllyh th~ col~
~y a n~g~tiYe pre~sure d~ r~n~al b~tw~en the opening~ 52 ~n~l B4 .
~ he ohroma~og;~ap~y p~oceclure ~y be co~enc~d by re~ov$ng c~lumn ho~ding a~se~ably 40 ~rom the ba~e 10 and inearting ~he collect~on vial 30 ~nto th~ ape~ture 22 in ~ a vial holding :~
a~3e~b1y 20. T~ aol~Dm hol~ing a~e~bly 40 L~ the~ return~d ~o -~
the base 10. ~ha ~olu~nn ~0 i3 in~er~ed ln~o 'c~ ape~tur~ 42 ln .
v~ g ~ P, v / l 4 ', ' 2~06~30 ~ a~
as3 th~ ¢olu~ holding a&sembly 40 an~ pol~ition~d ~4 'chat th~ low~r end o~ th~ colu~n extend~ lnto th~ ~ollqctlon vi~l 3 0 . An ~pprop~iat~ ~hro~atography ~ater~al mAy then bo intr4duced into the ~olu~n 60 u~inq the ~yringe 90, ~ other ~uitabl~ means.
I~ un~noo~porat~d nucleot~des ~re ~o b~ E~mo~ed ~roall DNA or ~NA ~ragmer~tE~, gQl c~rom~tog~ea~l~y r~aterial ~uch ~ a poly~acchar~d~ or polyacryla~ld~ a~r$rlg ~ ~olected de~e o~
~nts~n~l porosity, may ~e employed~ Th~ ple ~ontaining DN~ or ~NA ~ragments ~larg~ mvleculeB) and unlrlc~porat~d nu~leotlde~ls t~malle~ ~oleculQ~) may be lntroducQd ~nto th~ top o~ colu~n 40 u~ng a ~uitable p~pstting dev~c~. C~pllla~y a~tion d~aw~ tha ~plo into the upper po~tion o~ the ohroma~ ph5,~ ~aterisl, ~ .
l . e., between th~ "bead~" aompr~ng the mato~lal ag 5ho~ in ~:
Figu~ 2. ~e~ar~l~ly, i~ a 1 ml colu~nn is ~ployed, abou~ 10 -~0 Ul~ pre~r~bly 50 Ul~ and no mor~ ~ha~l ~bou~ 200 ml~ o~ s~mple `.
m~y b~ lntro~uc~d. With th~ column ~hU~ prepared, the ~y~inge gO
~ay ~e atta~had with t~e plu~3sJer ~ully extes~ded to the colual~n 20 The plungQr m~y thQn ba ~ir~ly depre~d ~with or without uQe 0 ~he support a~e~bly 50) untll 'che ~ample i~ pu~hed th~ough the coltlmn into ~ha colleo~$on v~al 30. If the #uppo~t as~embly 50 ~ eJaployed, ~he ~upport a3~emb1y lo¢lcing mechanism can be actuated to re~ain th~ plunger ~n a ~ully depressad co~ld$tion.
As ~he ~a~pl~ proc~ed~ throu~h the ~ natography mat~rlal, ~he ~malle~ molucules, ~or sx~mp~Q~ unincorporated nucleotidas~ arQ
partl~ioned into th~ pore6 in the ~hro~atog~aphy ~at~rlal Whil~
~h~ la~ge ~olucu~e~, ~or examp~,e, DNA or RNA ~ragment~, ~re e~lud~d. ~h~ ~luent ~rom th~ ~olunn should b~ ~ubs~an~ally ee o~ un~n~orpor~d n~cle4tide~ ~
A 8$~ilar proc:edura ~ay be employed for aff~nlty Gh~o~ agraphy a~li~at~on6 ~Uch .9 hy~rldizatlon o~
Li~,G, .8 ~ , p ~0~688~
l~S~J.6~C ;
co~pl~menta~ rando o~ nuGl~ic acl~. For ~xal~pl~ to s~pa~a~e poly~enyl~t~d ~PSA ~rom ~wnpolye.de2lylat~8 ~NA, oligo~
~ellulo~e ~ay ~e ~3~p~0yed a~ a ch~o~Q2lto~aphy mater~al. Unde~
appropr~te bu~er oon~l~tions, the de6~red poly~enylatad RN~
will bind wit~ the ollgo td~)-c¢llulo~o ~ o~ ography ~ateri~l wh~ he nonpoly~lenyl~t~d ~ 1 be o~ed ~nto the ` ~:
~olle~:tion v~al 30. ~he poly~deslylated ~A oan b~ reoo~rer~d by a, s~con~ bur~r ~:ond$~on. Other ai~nity ch~o~noto5rap~y appli~tlon~ inolllde the p~$~ioa~10~ o~ speci~lc nucleio ~cid :.
~oguences, ~or ~xa~nple, ~ral geno~Rlo 4s~uences, by generstin ~:o~plo~Qntary ollgonucl~o~lde~.
Thu8, ~ app~rAtu~ at~d chro~ tegr~phy 2~ethod e~ployi~g a pn~umat~c pre~u~a dl~erential ha~ro b~n dlaclo~ed. Whil~
embodl~nt~ and appllca~onA o~ thi~ ~nvent~osl have bebn ahown ahd d~ ib~d, it would bQ ~ppar~nt to thoa~ akillea in th~ a~t ~t many more modl~i¢atlon~ are ~o~lbl~ ~lt~out depa~tlng ~ro the lnventlvQ concept~ herein. For exa~ple, although puri~icat~on o~ 4~A and ~NA ~rag~ent~ and sap~at~on o~
polyadenylat~ad from nonpolyadenylated R~As ha~ been d-~sc~osed, many oth~r ~:h~omo~og~aphy appli~ations would be pos~ he ~ ~ .
inv~nt~on, thore~o~e, i3 not to be ~e8tr~ct~d except ir the ~: -~pi~it o~ tha appended olalm~
'; 20~6 ~a~
~85~153 Ç ;C ~ ~ Q J~ 8 /, ~ .
~ e p~e~ent lnv~n~ion relate~ l;o an spparatu~ a~d ~aethodology ~or th2 ch~o~atograp~ o~ mate~ial~, and in parti~ula~, ohro~atogr~phy b~ ed on mol~au~ar ~ , a~inlty and the like ~ used, ~o~ exampl~, in t~ purlfiQation~ ~ep~ra~iDn or i~41atior~ o~ ~NA and RNA ~ragment~, pro~Qln~ an~ other mole~ul~s.
a. ~5~
Re~oving unlnoorpo~ated nucleotide~ from DNA and ~NA
P~ag~nt6, i~ola~ing ~NA fractlons, pu~i~y~n~ protQ~ns and oth2r maoro~ oulo~, ar~ important prooedures having a v~riety o~
applicAtions. ~n ~N~ and RNA ~ynth~8i~, unlnc~r~oratQd nualoo~ld~s mu~t o~ten bQ removed wh~n ~on~trua~iny nick-tran~lat~d prob~, RNA probea and end-labeled ollgonuoleotidee, ~3 woll ~ illed-in" DNA ~ragmcnt~ i~portant ~o s~parat~ the unlncorporated ~Qe-nuelao~ides from the lab~l prob~ a~ unln~oxporated lab~l may bind to the sQlid ~uppor~, re~ulting in una~cep~ably hlg~ level~ of background nol~e.
~olat~on o~ ~NA ~actions ~a~ b~ Qmploy~d in th~ ~epara~ion of, ~
~or exa~pl~, poly~denylat~d RNA ~o~ nonpoly~danylat~d RNA~. ThQ .
u~ or ohromatoyra~hy m~thods to isolato and identi~y prot~ln~
and oth~ maoromole~ules i6 anothe~ well ~nown applicat~on. -`
Current ch~o~tog~aphy methods, u~e~ part~ularly i~
c~nnectlon w~ DNA an~ 6yn~ssls, tnclud~ ion-~xchange ahrom~tog~aphy, ~eYe~al variation~ of ~1 ch~omatography and other~. Each has i`~9 own d~advantag~s. For exa~ , ion-ex~hange ~thods ~ui~e a n~ber o~ step3 which ~y ~e~lt in a si~lfic~nt invQstment o~ ~im~ an~, in th~ ca~ o~ rad~ol~bele~
' ~ '.'' DE~, 2~'8~ i6:~^3 ~ . 5/ 6 ~9~
~s/a~s ~ ~:
nuol~otid~ ~ r~ng, ex~en~ive han~l:Ln~ o~ ~adioa~tive ~aterlal.
conve~tion~l ~el~ch~o~atogr~phy "drip~' ~olumn~ a~e ~diou~
r~uiring ~im~ to both pou~ ~n~ run~ ~p~n ~olumn~, A ~arlation o~ th4 i'drip~' column, ~r~ ~o~ewh~t ~a~3t~, but r~k r~atlo~
~xpo8~r~ and Gonta~in tion in the ~e of ra~onucl~ot~de chromatography, and m~y yie~d l~ss rQ~la~le r~sult~
A~ ~lt~natlve ~h~o~atogra~y approach whiah avo$d~ th~ ~
a~or~ment~on~d di~ficultie~ wou~d th~r~o~e ~ dQ~lrAb~e.
~8L:==~ ` `
~ h~ prQ~nt inv~ntlon i~ ~$~ct~d to an apparatu~ and ~ethod ~or ~uri~y~ng, iso~atlnq and separating ~teri~ls u~ing gel ~h~o~ato~aphy. To that end, a chroma~ography mat~rial an~ a ~mple may bs load~d into ~ column and pneu~at1q pro3~ur~ applled to uxg~ the ~ampl~ thxough ~he chromatography Pia,terial, whe~eby portion~ o~ the ~ampla may b~ aollacted by th~ chro~atog~aphy mat~rial and o~hor port~on~ exclu~d. In on~ ~bodl~ent, a posltl~e pn~u~atlc pr~sure iB prov~d~d ~nd in ~ s~cond ~mbodi~ent a n~gat~e pr~u~e i~ appl~d. Additlonally, a ~ovel ~upport etrU~tur~ ~ay be employed ~o 9upport the colu~n during ch~omatography. ~he ~ample may thus be gu~cXly and ral~ably trea~d.
F~gure 1 ~ a~ explodQd p~pective ~i~w o~ an appa~atus con~truot~d in a~co~ance wlth ~ha px~s~nt ~nv~ntlon co~prlslng ~ ~
column, prQssu~ in~uclng me~, a collectlon vial and asso~t~d ~ :
~upport ~ru~tur~
~ igura ~ 1B a ¢xo~ ct$onal view o~ ths appar~tu~ o~
F1gur~ a lo~ded poBiti ~E~, 28 8a !~
~0~i880 ~s~a~c }~efh~r~ng ~o Fi~ures 1 and 2 "~ oh~o~toq~apAy app~atus ¢onstruql:ed in accordan~e ~lth th~ E~reRe~t is)vent~or~ oomprise~ a gener~lly dl~k shaped ba~e ~0 havinc~ a palr o~ retainQr 1~ and a gen~ra~ly cyl~2~d~cal rial ho~ding as~embly 20 mo~nted th~xeon.
CQnt~ally located iTI the vial hold~y aa~e~ly 20 i~ a c~lln~ al ~ha~bar 22 ~or su~por~cing a collectlon v~l 30, ~rt~
whlch the elu~nt fro~ the oolumr~ m~y b~ coll~ted. ~he t~al 30 may bo a. dac~pped ~ppendo~f tubQ 02 otber ~ultab~ collecti~n ~an~. RQ~O~ab1Y mounted to t~ ba~ lo, and slidaa~ly engaglng ~h4 ex~erlor w~ll oS the v141 holding a~embly 20, ~g ~ gener~lly cylindrlcal c~lumn ~uppo~t as~e~bly ~0. ~he column ~upp~t a~e~bly 40 ln~lude~ a ~n~ral ~pert~rQ 42 ~or~ed ln ~he gone~ally plan~r uppar ~ux~ace 44 ~hereo~. As ~hown ~n Figure a, .;
the ~upp~t a0~em~1y ~ay hava A re~ nt aolla~ 46, ~uoh as an "0~' ~lng or th~ a, p~itloned ci~4um~rentially ad~ac~t the apexture 42, and a oollar re~alner 48 adapted t~ retain the coll~r ~¢ ad~a¢~nt ~he aper~ure 420 Al~ernatlYely, ~ ~hown in F1gU~G 1, th~ ~ollar 46 and the retainer 48 ~y bs ell~i~ated, optionally, a gene~ally a~lindrlcal pre6~ur~ induciny mean~
support as~e~ly 50 may be re~ovably mounted on the base 10. ~e ~.-support as~embly ma~ aompr~se a ¢~ntral a~rturs S2 ~on~ed 1~ the ~-~
g~ne~ally plan~r ~pper ~ur~¢~ B4 ther~o~, and 1~ ~on~igure~ to 61idsably engage tha ~xterio~ w~ll o~ the ~olu~ ~upport ~s~e~bly 40. ~he ~p~rt~e S~ i~ pr~rably ax~lly al~gned with th~ :~
apertuxe 4~ in the colu~n suppost sk~ucture 40, w~tch ~tself i6 .:
pre~e~s~ly axially sligned wi~h the ch~ber 22 in th~ vial holdin~ a~8~b1y 20.
~", 28 8~ ^ 5 . P. 1/14 ~006~
P~
18~7.
Al~er~ati~ , a~ ~hown $n Fi~U~çl 1., the~ 3uppor~ a~6am~1y 50 mBy ~nclude ~n upp~ u~Sac~ 54 ha~ing ro aperture t~erein. The ~upport ~embly 50 ma~ be ~ur~hex pro~ided with a pai~e o~
loa)ciJ~g tab~ 5~ adap~e~ to engage the ro~21in~rs 12 on th~ ~a~e 10 to lnotc th~ suppozt a~oe~nbly 50 ih p~a~:a du~in~ U8~. Oth~
sui'c~ble locXing me~hani~ms, such ~ thre~ds, could al~o be e~apl~yed. The assemblie~ ~0, 4~ ~nd 50 ~ay l:~e ~ormod o~ a radlatlon shlelding ~ate~l~l or, pre~erab~y, are ~on~t~uot~d to ~lt ~curely inside a bet~ ~hi~ld d~ ~loa. Molded P1A8tia nlateri~l~ have ~Q~n ~ound ~ult~ble although o~r mat~rial~ may ~1BO ~e ~snploy~d .
Supported ~y tlle column holding as~e~bly 40 ~bove l~he ~lal 30 i~ a ~ub~tsntially ~ubul~ oh~omatog~aphy column ~0. ~h~
colu~n 60 may b~ about 1 ~1 $n sizé, h~ing ~ p~a~erred lnta~nal d~amet~r o~ abo~t 5~ and a p~e~e~r~d length o~ a~out lOO~m, anq ~o~pri~e~ open~ngo ~a and 64, r~3p~ct$~ely, at each ~nd the~o~
An annula~ lip ~6 ~y b~ prov$ded olrcum~eren~ially ~d~acent ~h~
upp~r op~ning 62, a~ 5hown in Flgu~a 1. The upp~r ope~ln~ 62 is adapt~d to re~ei~o ~ ~romatog~p~y ~aterlal 70 and a 6~ple 80 to be flltered. ~he low~r op~ning 54 ~s ~ a~sa of rsduGed cro~ ection ad~pted to pre~ent pa~sag~ of th~ chro~ato~ap~y mat~r~al 70 w~le pa~t~$ng p~ a of the ~amplQ 80. ~`-Add~tlonally, a ecr~n or ~ er ~8, co~pr~sin~, ~or example, gla~ wool, may be employ~ to ~ntaln the Ghro~atogra~hY ~at~ial :~
70 withln ~he colu~n ~o. Pre~ra~ o~ ~ tubular ~olu~n, the -int~rnal dia~t~ ~hould not exceed abou~ lo~ ~or ~o~
~hromoto~r~phy appll~a~on6, lest the su~oe t~ion o~ the sample ~ in~u~ficlent to ~r~v~nt ~farve~o~nce ~nd cons~guent lo~ o~ p~e~s~Q t~rou~h tho samplo. In~reasing ~he len~ og ~ha colu~n 60 ~hould ~nhanc~ ~h~ deg~ae o~ ~par~tlon.
DE~, 28 8g liv:~t , P. 2~ l ' , . t_, j ~00688 PA~
~8S~t Pr~e~ably, the co1umr~ should b~ ns 14~ ~han ~bout 60mm in 1ength.
Pneuma~ic pre~sure ~nducing me~lh~ 94, ill thi~ ~a~e a syriTIge~ raay ~IQ at~ached to the ~lpp~r oper~ng ~2 o~ the Gol-~mn 60. A~ ~hown ~n ~igu~ 1, th~ sy~in~e ~0 may include a ~e~1e~ oS
'ch~ad~ ~2 w~1ch engage the annu1aP lip 66 o~ ~h~ colu~ 60 to . ~etaln the colu~n ~0 and th~ ~y~ln~a 90 in mutu41 engay~ment. In ~ ~1r3t ~m~od~ment o~ the inven~c10n, the syr:Ln~ gO, ~v~ng t~e plung~r 94 wlthdrawn as ~hown ~2~ F~g-lrs 2, m~y be a~taohed ~o ~he ~olu20n 60 an~ a po~tive pnsu~atio press~re d1~ferential ~ppl~ed between the open~ng~ 62 a~d 64 o~ the ~olu~n ~0 ~y d~pree~in~ t~
plun~er. In tha~ cas~, the support a0~embly 50 ~ay b~ place~
o~er the ~u~port as~ly 40. A~ the asse~ly 50 ls lowe~ B
upper ~ur~ce 5~ nc~ ap~rtur~ 52 i~ provl~ed) ~ill contaa~ ~ha plung~r 94 and auto~at~ally d~re~s ~am~ until the bott~ o~ the ~3~ppo~ 4em~1y SO Jneets thu base 10. ~t ~}~at polnt the 3upport 21~8~1~11bly 50 ~ay be ~iated unt$1 the locking tab~ 54 ~t~ t~rith ~;
the r~ta~n~r~ 1~. r~ an ~p~rture 52 ~L~ pro~id~d 1~ tbo ~upport ~esembly 50, t~ pl~ng~ 94 wlll ~x'eend there~hrough and may ~
~anually depre~d. ~n an alt~rnati~re embod~ont, not shown, the prQ~sure indu~lng mean~ 90 may be attached to the lower open~n~
~4 o~ thq colu~n ~th~ ~olu~n may be re~nove~ fro~ the ~upE:or~ -~se~bly 40 ln ~hat ~:a~e) and t~e sampls drawrl throllyh th~ col~
~y a n~g~tiYe pre~sure d~ r~n~al b~tw~en the opening~ 52 ~n~l B4 .
~ he ohroma~og;~ap~y p~oceclure ~y be co~enc~d by re~ov$ng c~lumn ho~ding a~se~ably 40 ~rom the ba~e 10 and inearting ~he collect~on vial 30 ~nto th~ ape~ture 22 in ~ a vial holding :~
a~3e~b1y 20. T~ aol~Dm hol~ing a~e~bly 40 L~ the~ return~d ~o -~
the base 10. ~ha ~olu~nn ~0 i3 in~er~ed ln~o 'c~ ape~tur~ 42 ln .
v~ g ~ P, v / l 4 ', ' 2~06~30 ~ a~
as3 th~ ¢olu~ holding a&sembly 40 an~ pol~ition~d ~4 'chat th~ low~r end o~ th~ colu~n extend~ lnto th~ ~ollqctlon vi~l 3 0 . An ~pprop~iat~ ~hro~atography ~ater~al mAy then bo intr4duced into the ~olu~n 60 u~inq the ~yringe 90, ~ other ~uitabl~ means.
I~ un~noo~porat~d nucleot~des ~re ~o b~ E~mo~ed ~roall DNA or ~NA ~ragmer~tE~, gQl c~rom~tog~ea~l~y r~aterial ~uch ~ a poly~acchar~d~ or polyacryla~ld~ a~r$rlg ~ ~olected de~e o~
~nts~n~l porosity, may ~e employed~ Th~ ple ~ontaining DN~ or ~NA ~ragments ~larg~ mvleculeB) and unlrlc~porat~d nu~leotlde~ls t~malle~ ~oleculQ~) may be lntroducQd ~nto th~ top o~ colu~n 40 u~ng a ~uitable p~pstting dev~c~. C~pllla~y a~tion d~aw~ tha ~plo into the upper po~tion o~ the ohroma~ ph5,~ ~aterisl, ~ .
l . e., between th~ "bead~" aompr~ng the mato~lal ag 5ho~ in ~:
Figu~ 2. ~e~ar~l~ly, i~ a 1 ml colu~nn is ~ployed, abou~ 10 -~0 Ul~ pre~r~bly 50 Ul~ and no mor~ ~ha~l ~bou~ 200 ml~ o~ s~mple `.
m~y b~ lntro~uc~d. With th~ column ~hU~ prepared, the ~y~inge gO
~ay ~e atta~had with t~e plu~3sJer ~ully extes~ded to the colual~n 20 The plungQr m~y thQn ba ~ir~ly depre~d ~with or without uQe 0 ~he support a~e~bly 50) untll 'che ~ample i~ pu~hed th~ough the coltlmn into ~ha colleo~$on v~al 30. If the #uppo~t as~embly 50 ~ eJaployed, ~he ~upport a3~emb1y lo¢lcing mechanism can be actuated to re~ain th~ plunger ~n a ~ully depressad co~ld$tion.
As ~he ~a~pl~ proc~ed~ throu~h the ~ natography mat~rlal, ~he ~malle~ molucules, ~or sx~mp~Q~ unincorporated nucleotidas~ arQ
partl~ioned into th~ pore6 in the ~hro~atog~aphy ~at~rlal Whil~
~h~ la~ge ~olucu~e~, ~or examp~,e, DNA or RNA ~ragment~, ~re e~lud~d. ~h~ ~luent ~rom th~ ~olunn should b~ ~ubs~an~ally ee o~ un~n~orpor~d n~cle4tide~ ~
A 8$~ilar proc:edura ~ay be employed for aff~nlty Gh~o~ agraphy a~li~at~on6 ~Uch .9 hy~rldizatlon o~
Li~,G, .8 ~ , p ~0~688~
l~S~J.6~C ;
co~pl~menta~ rando o~ nuGl~ic acl~. For ~xal~pl~ to s~pa~a~e poly~enyl~t~d ~PSA ~rom ~wnpolye.de2lylat~8 ~NA, oligo~
~ellulo~e ~ay ~e ~3~p~0yed a~ a ch~o~Q2lto~aphy mater~al. Unde~
appropr~te bu~er oon~l~tions, the de6~red poly~enylatad RN~
will bind wit~ the ollgo td~)-c¢llulo~o ~ o~ ography ~ateri~l wh~ he nonpoly~lenyl~t~d ~ 1 be o~ed ~nto the ` ~:
~olle~:tion v~al 30. ~he poly~deslylated ~A oan b~ reoo~rer~d by a, s~con~ bur~r ~:ond$~on. Other ai~nity ch~o~noto5rap~y appli~tlon~ inolllde the p~$~ioa~10~ o~ speci~lc nucleio ~cid :.
~oguences, ~or ~xa~nple, ~ral geno~Rlo 4s~uences, by generstin ~:o~plo~Qntary ollgonucl~o~lde~.
Thu8, ~ app~rAtu~ at~d chro~ tegr~phy 2~ethod e~ployi~g a pn~umat~c pre~u~a dl~erential ha~ro b~n dlaclo~ed. Whil~
embodl~nt~ and appllca~onA o~ thi~ ~nvent~osl have bebn ahown ahd d~ ib~d, it would bQ ~ppar~nt to thoa~ akillea in th~ a~t ~t many more modl~i¢atlon~ are ~o~lbl~ ~lt~out depa~tlng ~ro the lnventlvQ concept~ herein. For exa~ple, although puri~icat~on o~ 4~A and ~NA ~rag~ent~ and sap~at~on o~
polyadenylat~ad from nonpolyadenylated R~As ha~ been d-~sc~osed, many oth~r ~:h~omo~og~aphy appli~ations would be pos~ he ~ ~ .
inv~nt~on, thore~o~e, i3 not to be ~e8tr~ct~d except ir the ~: -~pi~it o~ tha appended olalm~
Claims (23)
1. A push column comprising a column adapted to hold a chromatography material and a sample, and means for inducing a pneumatic pressure differential across said column of sufficient magnitude to urge said sample through said chromatography material.
2. The device set forth in claim 1 wherein said column has a volume of about 1 ml and said sample is at least about 10 ul.
3. The device set forth in claim 1 wherein said column has a volume of about 1 ml and said sample is about 50 ul.
4. The device set forth in claim 1 wherein said column is a tube having a diameter of about 5mm.
5. The device set forth in claim 1 wherein said column comprises an upper opening adapted for introduction of chromatography material and a sample into said column, and a lower opening including a filter to prevent egress of chromatography material while permitting passage of a sample.
6. The device set forth in claim 1 wherein said column comprises a pair of openings and wherein said pressure inducing means comprise a syringe attached to one of said column openings.
7. The device set forth in claim 1 wherein said column comprises a pair of openings and wherein said pressure inducing means comprises a syringe attached to one of said column openings adapted to impart a positive pneumatic pressure to said opening, whereby said sample is urged away from said opening.
8. The device set forth in claim 1 wherein said pressure inducing means comprises a syringe attached to one of said column openings adapted to impart a negative pneumatic pressure to said opening, whereby said sample is drawn toward said opening.
9. The device set forth in claim 1 further including a support structure comprising means for supporting said column and means for supporting a collection vial.
10. The device set forth in claim 1 further including a support structure comprising means for supporting said column, means for supporting said pressure inducing means and means for supporting a collection vial.
11. A chromatography apparatus comprising means for supporting a column having a chromatography material and a sample therein during application of a pneumatic pressure differential sufficient to urge said sample through said chromatography material, and means for supporting a collection vial below said column.
12. The device set forth in claim 11 further including means for supporting a pneumatic pressure inducing means in engagement with said column.
13. The device set forth in claim 12 wherein said column support means and said means for supporting said pneumatic pressure inducing means are configured to fit within a radiation shielding device.
14. The device set forth in claim 11 wherein said column support means comprises an opening having a resilient collar to slideably engage said column.
15. A chromatography method comprising the steps of introducing a sample into a column containing a chromatography material, said chromatography material being adapted to collect portions of said sample while excluding other portions, inducing a pneumatic pressure differential across said column sufficient to cause said sample to pass through said chromatography material, and collecting a selected portion of said sample.
16. The method set forth in claim 15 wherein said chromatography material is a resin adapted to absorb selected portions of a sample having a predetermined molecular size limit while excluding portions of said sample having a molecular size that exceeds said limit.
17. The method set forth in claim 16 wherein said sample comprises DNA or RNA material having radiolabelled nucleotides incorporated therein and free radiolabeled nucleotides, and wherein said free radiolabelled nucleotides are absorbed by said chromatography material and said DNA or RNA material is eluted from said column.
18. The method set forth in claim 15 wherein said chromatography material separates on the basis of affinity.
19. The method set forth in claim 15 wherein said sample comprises a nucleic acid sequence and wherein said chromatography material comrpises an oligonucleotide complimentary to said nucleic acid sequence.
20. The method set forth in claim 15 wherein said sample comprises polyadenylated RNA and nonpolyadenylated RNAs, wherein said chromatography material comprises oligo(dT)-cellulose, and wherein said polyadenylated RNA binds with said oligo(dT)-cellulose chromatography material and said nonpolyadenylated RNAs are eluted from said column.
21. A push column comprising:
a column adapted to hold a chromatography material and a sample, said column being substantially tubular and having lower and upper openings therein, said upper opening having an annular lip circumferentially adjacent thereto;
means for inducing a pneumatic pressure differential between said column openings, said pressure differential inducing means being threadably attached to said annular lip of said column and having plunger means to generate said pressure differential;
a collection vial positioned below the lower opening of said column; and means for supporting said column above said collection vial.
a column adapted to hold a chromatography material and a sample, said column being substantially tubular and having lower and upper openings therein, said upper opening having an annular lip circumferentially adjacent thereto;
means for inducing a pneumatic pressure differential between said column openings, said pressure differential inducing means being threadably attached to said annular lip of said column and having plunger means to generate said pressure differential;
a collection vial positioned below the lower opening of said column; and means for supporting said column above said collection vial.
22. A push column comprising:
a column adapted to hold a chromatography material and a sample, said column being substantially tubular and having a pair of lower and upper openings therein, said upper opening having an annular lip circumferentially adjacent thereto;
means for inducing a pneumatic pressure differential between said column openings, said pressure inducing means being threadably attached to said annular lip of said column and having a plunger means to generate said pressure differential;
a collection vial positioned below the lower opening of said column; and a support structure comprising:
a base;
a collection vial support assembly having a central chamber therein to support said vial;
a column support assembly adapted to slideably engage said vial support assembly, said column support assembly comprising an upper surface having an aperture therein adapted to support said column; and a pressure inducing means support assembly adapted to slideably engage said column support assembly, said pressure inducing means support assembly comprising an upper surface adapted to engage said plunger means.
a column adapted to hold a chromatography material and a sample, said column being substantially tubular and having a pair of lower and upper openings therein, said upper opening having an annular lip circumferentially adjacent thereto;
means for inducing a pneumatic pressure differential between said column openings, said pressure inducing means being threadably attached to said annular lip of said column and having a plunger means to generate said pressure differential;
a collection vial positioned below the lower opening of said column; and a support structure comprising:
a base;
a collection vial support assembly having a central chamber therein to support said vial;
a column support assembly adapted to slideably engage said vial support assembly, said column support assembly comprising an upper surface having an aperture therein adapted to support said column; and a pressure inducing means support assembly adapted to slideably engage said column support assembly, said pressure inducing means support assembly comprising an upper surface adapted to engage said plunger means.
23. The device of claim 22 further including a locking mechanism to lock said pressure inducing means support assembly to said base.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US29280889A | 1989-01-03 | 1989-01-03 | |
| US292,808 | 1989-01-03 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2006880A1 true CA2006880A1 (en) | 1990-07-03 |
Family
ID=23126286
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002006880A Abandoned CA2006880A1 (en) | 1989-01-03 | 1989-12-29 | Push column and chromatography method |
Country Status (3)
| Country | Link |
|---|---|
| US (4) | US5378359A (en) |
| JP (1) | JPH03197861A (en) |
| CA (1) | CA2006880A1 (en) |
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| EP2695656A1 (en) | 2012-08-09 | 2014-02-12 | F. Hoffmann-La Roche AG | Method and separation device for separating a filtrate from a sample liquid |
| WO2014071965A1 (en) | 2012-11-12 | 2014-05-15 | Christian-Albrechts-Universität Zu Kiel | Nucleic-acid binding compounds |
| DE202014105965U1 (en) * | 2014-12-10 | 2015-01-26 | Cell.Copedia GmbH | Syringe housing for pipetting a biological material with integrated membranes |
| CN106317166A (en) * | 2015-07-02 | 2017-01-11 | 北京中原领先科技有限公司 | Device for separating proteins by using high-rate filtration and chromatography mixed mechanism |
| GB201519565D0 (en) * | 2015-11-05 | 2015-12-23 | Alere San Diego Inc | Sample preparation device |
| KR20220056442A (en) * | 2020-10-28 | 2022-05-06 | 주식회사 시선바이오머티리얼스 | Method for Extracting Nucleic Acid Using Syringe-Column Extraction System |
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| US3493503A (en) * | 1967-05-19 | 1970-02-03 | Haematronics Inc | Method of producing a protein-free fluid |
| US3682315A (en) * | 1969-10-20 | 1972-08-08 | Wolfgang Haller | Cartridge type column for treatment of liquid streams and substrate enclosure therefor |
| CA947997A (en) * | 1970-12-07 | 1974-05-28 | Charles J. Filz | Centrifugal chromatography apparatus and system |
| US3902849A (en) * | 1971-08-19 | 1975-09-02 | Medi Physics Inc | Radioisotope and radiopharmaceutical generators |
| DE2256357A1 (en) * | 1972-11-17 | 1974-06-12 | Traub Gmbh | DEVICE FOR FEEDING MATERIAL BARS FOR A MACHINE TOOL, IN PARTICULAR AUTOMATIC LATHE |
| US4138474A (en) * | 1973-05-01 | 1979-02-06 | Wisconsin Alumni Research Foundation | Method and device for immunoassay |
| US3884802A (en) * | 1973-10-05 | 1975-05-20 | Packard Becker Bv | Liquid chromatography injection system |
| US4168147A (en) * | 1977-12-02 | 1979-09-18 | Isolab, Inc. | Method to determine a diagnostic indicator of blood sugar condition, and, a liquid chromatographic microcolumn therefor |
| US4214993A (en) * | 1978-04-03 | 1980-07-29 | E. I. Du Pont De Nemours And Company | Apparatus for separating fluids |
| SU1041925A1 (en) * | 1979-08-16 | 1983-09-15 | Специальное Конструкторское Бюро Газовой Хроматографии | Gas chromatograph |
| US4270921A (en) * | 1979-09-24 | 1981-06-02 | Graas Joseph E | Microchromatographic device and method for rapid determination of a desired substance |
| US4335872A (en) * | 1980-03-10 | 1982-06-22 | Caplis Michael E | Sample preparation support apparatus |
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| US4388272A (en) * | 1981-04-08 | 1983-06-14 | Northwestern University | Method and apparatus for precise control of vapor phase concentrations of volatile organics |
| US4416775A (en) * | 1981-12-14 | 1983-11-22 | Std Filter Company, Inc. | In-line filter and cartridge therefor |
| DE3207386A1 (en) * | 1982-03-02 | 1983-09-15 | Bodenseewerk Perkin-Elmer & Co GmbH, 7770 Überlingen | METHOD AND DEVICE FOR APPLYING A LIQUID SAMPLE TO A GAS CHROMATOGRAPHIC SEPARATION COLUMN DESIGNED AS A CAPILLARY COLUMN |
| US4476017A (en) * | 1983-06-03 | 1984-10-09 | Beckman Instruments, Inc. | Removable synthesis column |
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| US4732672A (en) * | 1987-03-26 | 1988-03-22 | Kiang Chih Hen | Universally applicable mounting apparatus for high pressure liquid chromatography columns |
| IT210344Z2 (en) * | 1987-05-25 | 1988-12-06 | Sta Te Spa | EQUIPMENT FOR THE MEASUREMENT OF THE SPEED OF ERITROSEDIMENTATION OF BLOOD THROUGH PIPETTES. |
| JPH01195362A (en) * | 1988-01-29 | 1989-08-07 | Shimadzu Corp | Sample injecting device |
| US5186839A (en) * | 1988-06-03 | 1993-02-16 | Manac Inc. | Syringe type column for chromatography |
| EP0393185B1 (en) * | 1988-06-03 | 1995-08-23 | Manac Incorporated | Syringe type column for chromatography |
-
1989
- 1989-12-29 CA CA002006880A patent/CA2006880A1/en not_active Abandoned
-
1990
- 1990-01-04 JP JP2000201A patent/JPH03197861A/en active Pending
-
1992
- 1992-01-30 US US07/827,995 patent/US5378359A/en not_active Expired - Lifetime
-
1993
- 1993-06-28 US US08/084,533 patent/US5378360A/en not_active Expired - Lifetime
- 1993-06-28 US US08/084,534 patent/US5336412A/en not_active Expired - Lifetime
-
1994
- 1994-04-25 US US08/232,713 patent/US5413708A/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| US5378360A (en) | 1995-01-03 |
| JPH03197861A (en) | 1991-08-29 |
| US5413708A (en) | 1995-05-09 |
| US5378359A (en) | 1995-01-03 |
| US5336412A (en) | 1994-08-09 |
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