CM Weak Ion - Sib Pauv Membrane Chromatography

SP Strong Cation Exchange Membrane Chromatography Khoom Taw Qhia 

 

2. Cov khoom zoo

Fast thiab Efficient High binding muaj peev xwm ua tiav ntawm cov nqi ntws mus txog 40 lub sij hawm sai dua li cov tshuaj resin chromatography. Piv nrog cov pa roj carbon monoxide-raws li chromatography, lub sijhawm txheej txheem siv daim nyias nyias chromatography tuaj yeem raug txo los ntawm 30-40 zaug. Qhov kev khiav hauj lwm txaus tus nqi yog nyob ib ncig ntawm 10 MV / min.

2.2 High Binding Efficiency Membrane chromatography nthuav tawm muaj peev xwm khi tau siab thiab cov nqi ntws siab ntawm qhov tsis tshua muaj siab poob, ua kom cov khoom siv biomolecules raug ntes hauv ib qho dhau los ntawm lub module.

2.3 Scalable thiab Flexible Tag nrho cov khoom ntawm daim nyias nyias chromatography tuaj yeem ua tau raws li cov kev xav tau sib txawv ntawm biomolecule purification, npog cov ntawv thov los ntawm cov txheej txheem kev loj hlob mus rau loj - teev ntau lawm. Cov qauv tshuaj ntsiav tso cai rau ib zaug - siv kev ua haujlwm lossis tuaj yeem ntxuav thiab rov siv dua tom qab cov txheej txheem tu kom raug.

2.4 Txhim kho Productivity Lub compact tsim minimizes chaw footprint. Los ntawm kev tshem tawm cov haujlwm xws li kev ntim khoom, tu, tu validation, thiab kem cia, kev ua haujlwm tuaj yeem pib tom qab qhov sib npaug yooj yim nrog qhov tsis tshua muaj. Txij li tsis tas yuav tsum muaj cov kab ntim khoom, tu, lossis khaws cia, cov nqi ua haujlwm tuaj yeem txo qis dua 50%.

 

 

3 Technical Parameters

3.1 Cov ntaub ntawv tsim qauv

	Lab scale	me me nplai	pilot scale	ntau lawm scale
membrane ntim	0.2ml ib	5ml ib	140 ml	5L
Membrane Support Structure	Polypropylene
membrane vaj tsev	Polypropylene
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% ethanolaqueoussolution

Kev sib piv qhia tau hais tias kev pabcuam lub neej ntawm SP membrane chromatography yog piv rau SP Sepharose 6FF.

Daim duab 1. Kev hloov pauv muaj peev xwm ntawm SP Membrane Chromatography Tom Qab Kev Siv Ntau Yam Kev Ntsuas nrog Lysozyme

Tsis tas li ntawd, peb tau soj ntsuam cov membrane chromatography rau nws cov kev ua haujlwm ntawm kev tshem tawm cov cell cell proteins thiab nucleic acids. Cov txiaj ntsig yog raws li nram 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	96.4	423	3.6	2.07	7	4.3	0.21
2	97.4	438	4.3	2.01	7	4.7	0.17
3	94.1	513	5.8	1.95	6	2.3	0.42
4	96.2	32	0.8	1.60	6	3.2	0.27
5	96.3	45	1.9	1.37	8	3.4	0.37
6	96.7	158	2.4	1.82	8	3.5	0.36
7	96.4	267	2.6	2.01	9	6	0.18
8	96.8	298	3.6	1.92	9	7	0.11
9	97.9	746	9.8	1.88	4	1.5	0.43
10	96.2	39	3.2	1.09	4	1.4	0.46

Table 1. DNA thiab Host Cell Protein Tshem Tawm Tus Nqi hauv CHO- Qhia IgG Khoom Siv SP Membrane Chromatography

Ib qho kev sim ua kom pom tau tias SP daim nyias nyias chromatography tuaj yeem tshem tawm cov kab mob zoo thaum tswj hwm tus nqi rov qab siab ntawm lub hom phiaj IgG.

Thaum muab piv rau Gudiling SP membrane chromatography nrog cov khoom lag luam txawv teb chaws, cov ntaub ntawv muaj peev xwm khi tau raws li hauv qab no:

Kev khicmuaj peev xwm (mg / ml)	Sartorius	PALL	Kev taw qhia
Lysozyme	25	32	29
Trypsin	22	27	25

Table 2. Binding muaj peev xwm ua tau zoo ntawm cov protein sib txawv hauv peb cov khoom thiab cov khoom sib tw

Kev ntsuam xyuas tag nrho qhia tau hais tias peb lub peev xwm khi tau muab piv rau cov khoom tuaj txawv teb chaws.

Daim duab 2. Binding Peev Xwm Kev Ntsuas ntawm SP Membrane Chromatography Module Siv Lysozyme raws li tus qauv

Piv nrog rau dynamic binding muaj peev xwm thiab cov khoom tuaj txawv teb chaws, nws tau lees paub tias feem ntau lub hom phiaj proteins yuav ntes tau thaum lysozyme eluted nyob rau hauv 190 mM NaCl.

Los ntawm cov xwm txheej sib txawv, peb pom tias cov elution profiles ntawm daim nyias nyias chromatography zoo ib yam li cov agarose gel, tab sis cov purity ntawm cov protein sib txawv heev ntawm cov ntsev sib txawv. Nyob rau hauv qhov tseeb R&D thiab ntau lawm, nws yog ib qho tsim nyog yuav tau txiav txim siab qhov zoo tshaj plaws elution tej yam kev mob kom tau siab -purity lub hom phiaj proteins.

 

4 Yam Kev Thov Kev Pabcuam

Kev tshem tawm DNA, kab mob, thiab host cell proteins

Kev ntes ntawm plasmids, kab mob, cov proteins, thiab purification ntawm oligonucleotides

Kev tshem tawm cov xim ntawm cov poov xab fermentation broth thiab siab - muaj peev xwm protein capture

 

5 Cov txheej txheem siv

5.1 Kev npaj thiab sib dhos khoom:

5.1.1:Nruab lub membrane chromatography module ntawm ÄKTA chromatography system raws li cov txheej txheem zoo ib yam li resin kab, kom ntseeg tau tias cov kev taw qhia ntws mus raws cov module cov xub thiab inlet orientation. Txuas lub module siv Luer xauv los yog clamp-style fittings rau kev ruaj ntseg kev koom ua ke rau hauv lub system.

5.1.2 Teem lub feed flow rate rau5-10 MV / minthiab degas lub module siv equilibration buffer, xyuas kom meej tias cov effluent tsis muaj cua npuas. Thaum degassed, txuas lub qhov hluav taws xob permeate mus rau chromatography system.

5.2: Ua ntej-Siv Kev Kho Mob:

5.2.1:

Teem lub feed flow rate rau5-10 MV / minthiab ua ntej- kho nrog0.5 M NaOH rau ntau tshaj 5 MV, kom ntseeg tau tias daim nyias nyias ncav cuag qhov sib npaug.

5.2.2: Nyob rau tib tus nqi ntws, ua ntej- kho nrog1 M NaCl rau ntau tshaj 5 MVlos xyuas kom meej tias daim nyias nyias mus txog qhov sib npaug.

5.3 Cov txheej txheem Chromatography

5.3.1 Teem lub feed flow rate rau 5-10 MV/min thiab sib npaug ntawm daim nyias nyias nrog equilibration tsis rau ntau tshaj 5 MV, kom txog rau thaum lub membrane mus txog tag nrho equilibrium.

5.3.2 Tom qab 0.22 μm ua ntej- pom, thauj cov qauv mus rau hauv daim nyias nyias kom txog thaum cov qauv thauj khoom tiav lossis lub peev xwm chromatographic khi tau mus txog.

5.3.3 Ntxuav nrog equilibration buffer rau ntau tshaj 10 MV kom txog rau thaum UV teeb liab rov qab mus rau lub hauv paus.

5.3.4 Siv gradient elution lossis stepwise elution raws li tus txheej txheem tsim, thiab sau cov feem ntau raws li xav tau.

5.4 CIP Post-Siv Kev Kho Mob - Membrane Chromatography Device CIP (Tsev nyob rau hauv qhov chaw)

5.4.1 Teem lub feed flow rate rau 5–10 MV/min thiab ua kev tu nrog 1 M NaOH rau ntau tshaj 10 MV, kom txog rau thaum UV teeb liab txo qis rau hauv qab lub hauv paus.

5.4.2 Tom qab ncig rau 30 feeb, hloov mus rau kev ntxuav dej kom txog thaum pH nce mus txog 7-8, tom qab ntawd txuas ntxiv ntxuav nrog 20% ​​ethanol kom txog thaum cov conductivity yuav luag tas li.

5.5 Membrane Chromatography Cia

Tom qab siv thiab ua tiav ntawm CIP, daim nyias nyias tuaj yeem raug tshem tawm thiab khaws cia los ntawm soaking hauv 20% ethanol, lossis nws tuaj yeem khaws cia rau hauv - kab ntawm chav tsev kub hauv cov tshuaj uas muaj 20% ethanol. Cov tshuaj 20% ethanol yuav tsum tau kuaj xyuas thiab hloov ib ntus.

 

6. Cov ntaub ntawv xaj xaj

Q Strong Anion Txauv Membrane Chromatography Capsule Lim

	Lab scale	me me nplai	pilot scale	ntau lawm scale
Khoom qauv	IEXQ0002ES	IEXQ0050ES	IEXQ0400 ES	IEXQ 5000 ES
Membrane Volume	0.2ml ib	5ml ib	400 ml	5L

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