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GE 71-5000-15 AD manual

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Table of contents for the manual

  • Page 1

    Instructions 71-5000-15 AD Pre-activated media GE Healthcare CNBr -activated Sepharose 4 Fast Flow Introduction Th e pre pa ra ti on an d us e of a f f ini ty c hro ma to gra phy m ed ia b y cou pl in g bio sp ec if i c li gan ds t o CNBr-a c ti vat ed m at ri ce s is a w ide ly u se d, su cce ss f ul an d we ll -d oc ume nte d technique. CNBr -act[...]

  • Page 2

    p. 2 1. Product description CNB r -a c ti vat ed S ep har os e 4 Fas t Fl ow i s a be ad -fo rm ed , hig hl y cro ss- lin ke d pre - ac t iva te d ma tr ix p rod uce d by r ea ct in g Se ph aro se 4 Fa s t Fl ow wi th cy an og en b ro mid e ( CNB r) . Th is co up li ng ma ke s th e me diu m mo re ri gi d whi ch in t ur n im pro ves t he p re ss ure[...]

  • Page 3

    p. 3 Ta b l e 1 . Medium charac teristic s. Me an p ar t ic le si ze 90 μ m Part icle siz e range 45– 1 65 μ m Be ad s t ru c ture Hi gh ly cr os s-l in ked 4 % ag aro se, s ph er ic al Linear flowrate Ba se m at r ix 150 –250 c m /h , 0.1 MPa (1 b ar ) , XK 50 /6 0 co lum n, b ed h ei gh t 25 cm Swe ll in g fac to r 4 –5 ml d rai ne d me d[...]

  • Page 4

    p. 4 In or de r to ret ain m ax im um b ind in g cap ac ity o f CNBr-a c ti vat ed S ep har os e 4 Fas t Fl ow p ri or t o cou pl ing t he l ig an d, us e co ld ( 0 – 4 ° C) s olu t ion s . Th e ti me in ter val b et w ee n was hi ng an d co upl in g mu st b e mi nim is ed ; t he refo re pre pa ra ti ons o f all r eq uir ed s olu t ion s pr io r[...]

  • Page 5

    p. 5 5. Was h awa y ex es s li gan d wi t h at l ea st 5 m ed ium v ol ume s of co up li ng b uf fe r. 6. Af te r cou pl in g, no n- re ac te d gro up s on t he m edi um s ho uld b e b lo cke d. T ran s fer t he m edi um to 0 .1 M T ris -H Cl bu f fe r pH 8. 0 or 1 M e th ano la mi ne pH 8 .0. Le t it s ta nd fo r 2 ho ur s. 7 . Was h th e cou pl e[...]

  • Page 6

    p. 6 Lar ge sc al e col umns • BP G TM var ia bl e be d, g la ss co lu mn s. In ne r di ame te rs f rom 10 0 –450 m m, be d vol um es f rom 2.4 – 131 lit res ; be d he ig ht ma x 83 cm . • CHRO MAFLO W TM var ia bl e be d col um ns . Inn er d iam et er s fro m 40 0 – 60 0 mm . 3.2 Packing lab-scale columns 1. As se mb le t he co lu mn ( a[...]

  • Page 7

    p. 7 7 . Wh en t he b ed h as s tab il ize d, c lo se t he b ot tom o ut le t an d st op t he p ump . 8. If u sin g a pa cki ng r es er vo ir, dis con ne ct t he r es er vo ir a nd f i t th e ad apt or to th e co lum n. 9. Wi th t he a da pto r inl et d is con ne c ted , pu sh d own t he a da pto r ap pro xim at el y 2 mm in to th e be d, a ll ow i[...]

  • Page 8

    p. 8 Th e cal cul ate d pl at e numb er w ill v ary d ep end ing o n th e tes t co nd iti ons and i t sho uld t he ref or e be us ed a s a ref er enc e val ue on ly. I t is a ls o imp o r tan t th at co nd it io ns a nd e qui pm en t are ke pt co ns ta nt so t ha t re sul t s are co mp ar abl e. Ch an ge s in s ol ute, s ol ven t, e lu ent , sa mp [...]

  • Page 9

    p. 9 Th e red uce d pl at e he igh t is ca lcu la te d : HET P/d wh ere d i s th e dia me ter o f th e be ad . As a g ui de lin e, a va lue o f <3 is n or ma ll y acceptable . Th e pe ak s hou ld b e sy mme tr ic al, a nd t he a sym me tr y fa ct or a s clo se a s po ss ibl e to 1 ( va lue s b et we en 0. 8 – 1. 5 ar e usu al ly a ccep tab le [...]

  • Page 10

    p. 10 5. Cleaning, Sanitization and Storage For b es t p er fo r man ce of co up le d CNB r -a c ti va ted S ep ha ros e 4 Fas t Fl ow o ver a lo ng wo rk in g li fe, fol lo w th e ge ner al pr oce du res d es cr ib ed b el ow. In al l cas es , we re co mme nd t es ti ng t he p ro ced ure s at s ma ll s cal e f ir st . Equilibration After packing, [...]

  • Page 11

    p. 11 or Was h th e col umn w it h 3– c ol umn v ol ume s of 70 % et han ol . Was h im me di ate ly w it h at l ea s t 5 col umn vol um es of s te ri le f i lte re d bin di ng b uf f er. Sanitization San it iz at io n in ac ti va tes m ic rob ia l cont am ina nt s in t he p ac ked c olu mn a nd rel at ed e qui pm ent . A s pe ci f ic sa ni ti za [...]

  • Page 12

    BioProcess, BPG, CHROMAFLOW, Drop Design, Sephar ose, and Tricorn are trademarks of GE Healthcare companies. GE, imagination at work and GE monogram are trademarks of General Electric Company. All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. GE Healthcare reserves t[...]