Q Strong Anion Membrane Chromatography

 

2. Cov khoom zoo

2.1 Ceev thiab ua haujlwm tau zoo - ua tiav cov peev txheej siab ntawm cov nqi ntws mus txog 40 npaug sai dua li cov khoom siv niaj hnub. Piv nrog rau cov khoom ntim - txaj chromatography, lub sijhawm txheej txheem siv daim nyias nyias chromatography yog txo los ntawm 30–40 zaug. Qhov kev khiav hauj lwm txaus tus nqi yog 10 MV / min.

2.2 High binding efficiency - membrane chromatography nthuav tawm lub peev xwm thauj khoom siab thiab cov nqi ntws siab nyob rau hauv qhov tsis tshua muaj siab poob qis, tso cai rau cov khoom siv biomolecules raug ntes hauv ib zaug.

2.3 Scalable thiab saj zawg zog - tag nrho series ntawm membrane chromatography cov khoom tuaj yeem ua tau raws li cov kev xav tau sib txawv ntawm biomacromolecule purification, npog txhua theem ntawm txheej txheem kev loj hlob mus rau loj - teev ntau lawm. Cov tshuaj ntsiav - hom kev tsim qauv txhawb nqa ob qho tib si - siv ua haujlwm thiab rov siv dua tom qab ntxuav.

2.4 Txhim kho cov khoom tsim tau zoo - Cov qauv tsim kom txo qis qhov chaw hneev taw. Los ntawm kev tshem tawm cov kab ntim khoom, tu, tu validation, thiab kem cia kev khiav hauj lwm, cov txheej txheem yuav tsum tau tsawg tsis rau equilibration thiab yuav ua tau ncaj qha yam tsis muaj kab npaj. Cov nqi ua haujlwm tuaj yeem txo qis txog 50%.

 

3. Technical parameter

3.1 Cov khoom siv zoo

	Lab scale	me me nplai	pilot scale	ntau lawm scale
membrane ntim	0.2ml ib	5ml ib	140 ml	5L
Membrane kev them nyiaj yug qauv:	polypropylene (PP)
membrane vaj tsev	polypropylene (PP)
O lub nplhaib	silicone

 

3.2 Cov yam ntxwv ua haujlwm

	Lab scale	me me nplai	pilot scale	ntau lawm scale
membrane ntim	0.2ml ib	5ml ib	400 ml	5L
Pom zoo txaus tus nqi	1-6 ml / min	25-150ml / min	2-12L / min	25-150L / min
Ua haujlwm kub siab tshaj plaws	35 degree
Kev ua haujlwm siab tshaj plaws	3bar (25 degree)
Qhov siab tshaj qhov sib txawv	3bar (25 degree)
Cia tej yam kev mob:	20% ethanol aqueous tov

 

Kev pabcuam lub neej ntawm Q membrane chromatography yog piv rau agarose gel chromatography.

 

 

Daim duab 1. Kev hloov pauv hauv daim nyias nyias chromatography loading peev tom qab siv ntau yam kev soj ntsuam los ntawm BSA

Tsis tas li ntawd, peb tau soj ntsuam qhov kev tshem tawm ntawm daim nyias nyias chromatography rau cov tswv tsev cell proteins thiab nucleic acids. Cov txiaj ntsig tau raws li hauv qab no.

	DNA				HCP
	IgG rov qab	Cov ntsiab lus (pg / mg ntawm IgG) los ntawm RT PCR		Kev tshem tawm Factor	Cov ntsiab lus (ng / mg ntawm IgG) los ntawm Elisa		Kev tshem tawm Factor
Khiav	%	Ua ntej Q Membrane	Tom qab Q Membrane	Log	Ua ntej Q Membrane	Tom qab Q Membrane	Log
1	96.7	423	1.2	2.55	7	4.8	0.16
2	97.4	438	1.3	2.53	7	4.9	0.15
3	94.7	513	1.3	2.60	6	1.9	0.50
4	95	32	0.9	1.55	6	4.2	0.15
5	96.3	45	1.1	1.61	8	5.2	0.19
6	96.5	158	0.7	2.35	8	6.2	0.11
7	96.4	267	1.9	2.15	9	7.2	0.10
8	96.8	298	2.3	2.11	9	8.2	0.04
9	97.1	746	1.5	2.70	4	2	0.30
10	96.6	39	1	1.59	4	1.5	0.43

Table 1. Tshem tawm cov nqi ntawm DNA thiab host cell proteins hauv CHO-qhia IgG khoom siv Q membrane chromatography

 

Ib qho kev sim ua kom pom tau tias Q membrane chromatography tuaj yeem tshem tawm cov impurities zoo thaum tuav lub hom phiaj siab IgG rov qab.

Tsis tas li ntawd, peb piv Gudiling membrane chromatography nrog cov khoom lag luam, thiab cov ntaub ntawv muaj peev xwm thauj khoom yog raws li hauv qab no:

Loading peev (mg / ml)	Sartorius	PALL	Kev taw qhia
BSA	29	60	45
SOD	25	40	35
Protein A	27	45	40
Trypsin	30	56	44

Table 2. Chaw thau khoom kev ua tau zoo ntawm cov protein sib txawv ntawm peb cov khoom thiab cov khoom sib tw

Kev ntsuam xyuas tag nrho qhia tau hais tias peb lub peev xwm thauj khoom yog piv rau cov khoom siv txawv teb chaws,

Daim duab 2.Siv lub Q membrane chromatography module, BSA tau siv los ua tus qauv protein los ntsuas lub peev xwm khi. Lub dynamic binding muaj peev xwm tau muab piv nrog cov khoom ntawm imported. Cov txiaj ntsig tau pom tias feem ntau ntawm cov hom phiaj protein tuaj yeem raug ntes thiab eluted nrog150 mM NaClthaum siv BSA ua cov qauv protein.

Los ntawm cov xwm txheej sib txawv, peb pom tias tus cwj pwm elution ntawm daim nyias nyias chromatography zoo ib yam li ntawm agarose gel chromatography. Lub purity ntawm cov proteins eluted ntawm txawv ntsev concentration nws txawv ho. Hauv kev tsim thiab tsim cov txheej txheem tiag tiag, nws yog ib qho tsim nyog yuav tsum txiav txim siab ntau qhov kev pom zoo elution nyob rau hauv thiaj li yuav tau txais lub hom phiaj protein nrog siab purity.

 

4. Cov ntaub ntawv thov ib txwm

Kev tshem tawm DNA, kab mob, host cell proteins (HCP), thiab endotoxins

Kev ntes ntawm plasmids, kab mob, thiab cov proteins, nrog rau kev ua kom huv ntawm oligonucleotides

 

5. Cov txheej txheem ua haujlwm

5.1: Kev npaj thiab sib dhos:

5.1.1: AKTA chromatography system: Nruab lub membrane chromatography module nyob rau hauv ib tug yam ntxwv zoo ib yam li packed txhua. Xyuas kom meej tias tus xub taw qhia ntawm lub module yog raws li cov kev taw qhia feed flow. Txuas lub kaw lus siv Luer txuas lossis Tri-Clamp fittings.

5.1.2 Teem lub inlet flow rate rau 5-10 MV / min thiab siv qhov sib npaug tsis sib npaug kom tshem tawm cov huab cua ntawm lub cev. Tom qab xyuas kom meej tias tsis muaj cua npuas hauv qhov hluav taws xob, txuas lub qhov hluav taws xob permeate mus rau chromatography system.

5.2: Ua ntej- siv Kev Kho Mob:

5.2.1:

Set the inlet flow rate to 5–10 MV/min and perform pre-treatment with 0.5 M NaOH at >5 MV los xyuas kom meej tias daim nyias nyias mus txog qhov sib npaug.

5.2.2:

At the same flow rate, perform pre-treatment with 1 M NaCl at >5 MV los xyuas kom meej tias daim nyias nyias mus txog qhov sib npaug.

5.3 Cov txheej txheem Chromatography

5.3.1 Set the feed flow rate to 5–10 MV/min and perform pre-treatment with equilibration buffer at >5 MV mus txog rau thaum daim nyias nyias mus txog qhov sib npaug.

5.3.2 Tom qab cov qauv ua ntej- lim los ntawm 0.22 μm lim, thauj nws mus rau hauv kem kom txog thaum tag nrho cov qauv siv los yog chromatography binding muaj peev xwm ncav cuag.

5.3.3 Flush with equilibration buffer at >10 MV mus txog rau thaum lub teeb liab UV poob rau hauv paus.

5.3.4 Ua gradient lossis linear elution raws li tus qauv tsim, thiab sau cov feem raws li xav tau.

5.4 CIP Post- siv Kev Kho Mob - Membrane Chromatography Device CIP

5.4.1 Set the feed flow rate to 5–10 MV/min and treat with 1 M NaOH at >10 MV kom txog rau thaum lub teeb liab UV poob qis hauv qab lub hauv paus.

5.4.2 Tom qab ncig rau 30 feeb, hloov mus rau dej ntxuav kom txog thaum pH nce mus txog 7-8, ces hloov mus rau 20% ethanol thiab ntxuav txuas ntxiv mus kom txog thaum lub conductivity stabilizes.

5.5, Membrane Chromatography Cia:

Tom qab siv thiab ua tiav ntawm CIP, lub membrane module tuaj yeem muab tshem tawm thiab khaws cia hauv 20% ethanol, lossis khaws cia hauv online ntawm chav tsev kub hauv 20% ethanol. Cov tshuaj ethanol yuav tsum tau hloov ib ntus thiab tshuaj xyuas.

 

6. Cov ntaub ntawv xaj xaj

Q Strong Anion-Exchange Membrane Chromatography Capsule Filter

	Lab scale	me me nplai	pilot scale	ntau lawm scale
Yam khoom Model	IEXQ0002ES	IEXQ0050ES	IEXQ0400 ES	IEXQ 5000 ES
membrane ntim	0.2ml ib	5ml ib	400 ml	5L

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