SP Strong Cation Txauv Membrane Chromatography

Taw qhia rau CM Weak Ion-Kev Sib Pauv Membrane Chromatography Cov Khoom

 

1. Txheej txheem cej luam

CM tsis muaj zog cation- pauv chromatography yog ib qho kev ua kom huv si uas cov carboxymethyl pawg tau sib koom ua ke rau hauv daim nyias nyias los tsim cov qauv ua haujlwm. Kev sib cais yog ua tiav raws li qhov sib txawv ntawm cov khoom siv hluav taws xob thiab cov nqi them nqi ntawm ntau yam biomolecules. Txij li feem ntau cov macromolecules lom muaj carboxyl lossis hydroxyl pawg, lawv cov yam ntxwv thiab qhov loj tuaj yeem hloov kho los ntawm kev hloov pH tus nqi ntawm cov tshuaj tsis haum. Tom qab cov biomolecules khi rau daim nyias nyias module nqa cov nqi sib txawv, elution yog ua los ntawm kev hloov lub zog ionic lossis pH ntawm lub sijhawm txawb. Molecules uas tsis muaj zog binding affinity yog eluted ua ntej, thaum cov uas muaj zog binding affinity yog eluted tom qab, yog li ua tau zoo sib cais.

 

2. Cov khoom zoo

2.1 Ceev thiab ua haujlwm tau zoo: Muaj peev xwm sib khi siab tuaj yeem ua tiav ntawm tus nqi ntws mus txog 40 npaug sai dua li cov pa roj carbon monoxide -raws li chromatography. Piv nrog rau cov khoom ntim ib txwm muaj - txaj chromatography, daim nyias nyias chromatography tuaj yeem txo cov sijhawm ua haujlwm los ntawm 30-40 zaug. Qhov kev khiav hauj lwm txaus tus nqi yog 10 MV / min.

2.2 High binding efficiency: Membrane chromatography qhia tau hais tias muaj peev xwm loading siab thiab siab txaus tus nqi nyob rau hauv uas tsis tshua muaj siab poob tej yam kev mob, enabling nqi biomolecules yuav tsum tau ntes nyob rau hauv ib tug dhau los ntawm lub module.

2.3 Scalable thiab saj zawg zog: Ua tiav ntau yam ntawm cov khoom siv membrane chromatography tuaj yeem ua tau raws li cov kev xav tau ntawm biomacromolecule, suav nrog cov theem los ntawm kev txhim kho mus rau loj - nplai tsim. Cov qauv tshuaj ntsiav tso cai rau ib zaug - siv daim ntawv thov lossis tuaj yeem ntxuav thiab rov siv dua raws li qhov xav tau.

2.4 Ntxiv cov khoom tsim tau: 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 cov kauj ruam, kev ua tiav tuaj yeem pib tom qab sib npaug nrog qhov ntim me me ntawm qhov tsis sib xws. Tsis tas yuav tsum tau ntim cov kab ke, tu, lossis khaws cia, cov nqi ua haujlwm tuaj yeem txo qis txog li 50%.

 

3 Technical Parameters

3.1 Cov ntaub ntawv tsim qauv

	Laboratory nplai	me me nplai	Pilot nplai	Ntau lawm nplai
Membrane ntim	0.2ml ib	5ml ib	140 ml	5L
Membrane kev them nyiaj yug qauv	Polypropylene
Membrane vaj tse	Polypropylene
O lub nplhaib	Silicone roj hmab

3.2 Cov yam ntxwv ua haujlwm

	Laboratory nplai	me - nplai	Pilot - scale	Ntau lawm nplai
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

Piv nrog rau cov pa xov xwm, kev pab cuam lub neej ntawm CM membrane chromatography yog piv rau CM agarose gel 6FF.

Daim duab 1. Kev hloov pauv ntawm lub peev xwm thauj khoom ntawm CM membrane chromatography tom qab siv ntau yam hauv kev kuaj lysozyme.

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

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

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

	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	6.9	1.79	7	6.1	0.06
2	97.4	438	5.8	1.88	7	4.8	0.16
3	94.7	513	9.6	1.73	6	3.6	0.22
4	95	32	4.6	0.84	6	4.7	0.11
5	96.3	45	4.6	0.99	8	5.1	0.20
6	96.5	158	8.2	1.28	8	4.6	0.24
7	96.4	267	6.5	1.61	9	6.8	0.12
8	96.8	298	5	1.78	9	7.4	0.09
9	97.1	746	5.6	2.12	4	3.5	0.06
10	96.6	39	5	0.89	4	3.9	0.01

Siv Gudiling CM daim nyias nyias chromatography piv nrog rau lwm hom, cov ntaub ntawv muaj peev xwm thauj khoom tau qhia hauv qab no.

Loading peev (mg / mL)	Jingbiao 1	Kev taw qhia
Lysozyme	18	23
Trypsin	17	20

Table 2. Kev thauj khoom ntawm cov protein sib txawv hauv peb cov khoom piv nrog cov khoom sib tw

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

Daim duab 2. Kev ntsuas lub peev xwm ntawm CM membrane chromatography module siv lysozyme ua cov protein ntau.

Lub peev xwm thauj khoom dynamic kuj tau muab piv nrog cov khoom tuaj txawv teb chaws. Kev txheeb xyuas pom tau tias lysozyme tuaj yeem raug tshem tawm siv 160 mM NaCl, uas tso cai rau kev ntes cov protein ntau lub hom phiaj.

Los ntawm kev ua kom zoo ntawm cov xwm txheej sib txawv, peb pom tias daim nyias nyias chromatography nthuav tawm tus cwj pwm elution zoo ib yam li agarose gel chromatography, qhov twg cov protein purity txawv heev nyob rau hauv cov ntsev sib txawv. Hauv kev ua tau zoo R&D thiab kev tsim khoom, nws yog ib qho tsim nyog los txiav txim siab qhov sib npaug ntawm cov ntsiab lus kom tau txais qib siab -purity phiaj protein.

 

4. Cov ntawv thov ib txwm

• Tshem tawm cov DNA, cov kab mob, thiab cov cell proteins
• Kev ntes cov plasmids, kab mob, thiab cov proteins, nrog rau kev ua kom huv ntawm oligonucleotides
• Decolorization ntawm poov xab fermentation broth thiab siab -muaj peev xwm protein capture

 

5. Kev khiav hauj lwm txheej txheem / Workflow

5.1: Cov cuab yeej npaj thiab sib dhos:

5.1.1: Nruab lub membrane chromatography module ntawm AKTA chromatography system Cov txheej txheem kev teeb tsa zoo ib yam li ntim - txaj chromatography media; xyuas kom meej tias cov kev taw qhia ntws raws li cov xub cim rau ntawm lub module. Lub module tuaj yeem txuas nrog siv Luer connectors lossis clamp connectors.

5.1.2 Teem lub inlet flow rate ntawm 5-10 MV / min thiab siv qhov sib npaug tsis sib npaug los ntxuav cov huab cua los ntawm lub module. Txuas ntxiv mus kom txog thaum tsis muaj npuas nyob ntawm qhov hluav taws xob, ces txuas lub qhov hluav taws xob permeate mus rau qhov system chromatography.

5.2.: Ua ntej- siv kev npaj

5.2.1: Teem lub inlet flow rate ntawm 5-10 MV / min thiab ua pretreatment siv 0.5 M NaOH rau ntau tshaj 5 MV los xyuas kom meej cov membrane ncav cuag equilibrium.

5.2.2: Nyob rau hauv tib lub ndlwg tus nqi, ua ntej- kho nrog 1 M NaCl rau ntau tshaj 5 MV los xyuas kom meej cov membrane ncav cuag qhov sib npaug.

5.3. Chromatography txheej txheem

5.3.1 Teem lub inlet flow rate rau 5–10 MV/min thiab ua ntej- kho nrog equilibration buffer rau ntau tshaj 5 MV kom txog rau thaum lub membrane mus txog qhov sib npaug.

5.3.2 Tom qab 0.22 μm ua ntej- pom, thauj cov qauv mus rau hauv kem thiab txuas ntxiv mus kom txog thaum tag nrho cov qauv tau siv los yog chromatography loading muaj peev xwm mus txog.

5.3.3 Ntxuav nrog equilibration buffer rau ntau tshaj 10 MV kom txog rau thaum UV nqi txo mus rau hauv paus.

5.3.4 Raws li tus txheej txheem tsim, siv gradient elution lossis linear elution system, thiab sau cov feem hauv ntu raws li xav tau.

5.4 CIP Post-siv kev kho mob – CIP (Huv-hauv-Place) rau membrane chromatography system.

5.4.1 Teem lub inlet flow rate rau 5–10 MV/min thiab kho nrog 1 M NaOH rau ntau tshaj 10 MV kom txog thaum tus nqi UV poob qis hauv qab lub hauv paus.

5.4.2 Tom qab 30 feeb ntawm kev ntxuav, yaug nrog dej kom txog thaum pH nyob nruab nrab ntawm 7 thiab 8, ces hloov mus rau 20% ethanol thiab txuas ntxiv tu kom txog thaum cov conductivity tseem yuav luag tsis hloov.

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 kev ntub dej hauv 20% ethanol, lossis khaws cia hauv online ntawm chav tsev kub hauv cov tshuaj uas muaj 20% ethanol. Cov tshuaj 20% ethanol yuav tsum tau kuaj xyuas thiab hloov tsis tu ncua.

 

6, Order information

CM - hom tsis muaj zog cation txauv membrane chromatography capsule lim

	Laboratory nplai	Me me	Pilot nplai	Ntau lawm nplai
Qauv	IEXCM0002ES	IEXCM0050ES	IEXCM0400 ES	IEXCM5000ES
Membrane cheeb tsam	0.2ml ib	5ml ib	400 ml	5L

Cim npe nrov: sp muaj zog cation txauv membrane chromatography, Tuam Tshoj, lwm tus neeg, manufacturers, hoobkas, lag luam wholesale, bulk, Tshuag, dawb qauv