Bio-Rad Bio-Scale Mini CHT 40 µm Cartridges User Manual
Bio-Scale™Mini CHT
™
Ceramic Hydroxyapatite
Cartridges, 5 ml
Instruction Manual
Catalog # 732-4322
732-4324
732-4332
732-4334
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5.2 Wash Alternatives ..................20
5.3 Autoclaving ............................20
5.4 Storage ..................................20
Section 6 ..Technical Support.............................21
Section 7 ..Ordering Information.........................22
Section 8 ..References .......................................24
The Bio-Scale Mini CHT cartridges are packed with
Type I or Type II ceramic hydroxyapatite supports.
These supports are based on hydroxyapatite, a
form of calcium phosphate used in chromatographic
separations of biomolecules. CHT ceramic
hydroxyapatite is a spherical, macroporous form of
hydroxyapatite. Unlike most other chromotagraphic
absorbents, CHT is both the ligand and the support
matrix. CHT Type I has a higher protein binding
capacity and greater capacity for acidic proteins.
CHT Type II has a lower protein binding capacity
but provides better resolution of nucleic acids and
certain other proteins. The Type II material also has a
very low affinity for albumin and is especially suitable
for the purification of many species types and classes
of immunoglobulins. Applications of hydroxyapatite
chromatography include the purification of different
subclasses of monoclonal and polyclonal antibodies,
antibodies that differ in light chain composition,
antibody fragments, isozymes, supercoiled DNA from
linear duplexes, and single-stranded from double
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Table 2. Product Description
TTyyppee II TTyyppee IIII
FFuunnccttiioonnaall ggrroouuppss
Ca
2+
, PO4, OH Ca2+, PO4, OH
OObbsseerrvveedd ddyynnaammiicc bbiinnddiinngg ccaappaacciittyy
llyyssoozzyymmee ((LLyyss))
≥ 25 mg Lys/g CHT ≥ 12.5 mg Lys/g CHT
NNoommiinnaall ppoorree ddiiaammeetteerr
600–800 Å 800–1,000 Å
MMaaxxiimmuumm bbaacckkpprreessssuurree
100 bar (1,500 psi) 100 bar (1,500 psi)
N
Noommiinnaall mmeeaann ppaarrttiiccllee ssiizzee
20 ± 2, 40 ± 4, and 80 ± 8 µm
BBuullkk ddeennssiittyy
0.63 g/ml 0.63 g/ml
OObbsseerrvveedd ddyynnaammiicc bbiinnddiinngg ccaappaacciittyy
IIggGG
25–60 mg IgG/ml CHT* 15–25 mg IgG/ml CHT*
TTyyppiiccaall lliinneeaarr fflloow
w rraattee rraannggee
50–1,000 cm/hr
ppHH ssttaabbiilliittyy
6.5–14
BBaassee ssttaabbiilliittyy
at least 21 months in 1 N NaOH
RReeggeenneerraattiioonn
500 mM sodium phosphate, pH 7
1,000 mM trisodium phosphate, pH 11–12
AAuuttooccllaavvaabbiilliittyy ((bbuullkk))
121°C, 20 min in phosphate buffer, pH 7
SSaanniittiizzaattiioonn
1–2 N NaOH
RRe
eccoommmmeennddeedd ccoolluummnn ssttoorraaggee
0.1 N NaOH
SShheellff lliiffee ((ddrryy,, uunnuusseedd mmaatteerriiaall))
85 months stored dry, sealed, and at room
temperature
* 40 µm particles, 300 cm/hr, 5 mM sodium phosphate, pH 6.5
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