Biomedical Engineering Reference
In-Depth Information
Table 3.8.
Buffers for elution in immunoaffinity chromatography
Buffer
pH
pH change
0.1 M glycine-HCl
1.5 - 2.8
0.1 M glycine-HCl, 0.5 M NaCl
2.5
0.1 M Na acetate, 0.15 M NaCl, HCl
1
0.1 M Na citrate/phosphate
3.0 - 3.5
0.1 M acetic acid/formic acid
2.2
0.5 M acetic acid
1 M propionic acid
0.15 M NaCl/NH
4
OH
11.0
0.1 M triethylamine
11.5
Chaotropic elution
2.5 M KSCN
Unbuffered
0.5 - 3 M NaSCN
Unbuffered
3 M guanidinium hydrochloride
Unbuffered
6 M urea
Unbuffered
10% 1,4-dioxane (v/v)
Unbuffered
80% ethylenglycol (v/v)
Unbuffered
High ionic strength
6.8 M NaCl
Unbuffered
4 M KI, 20 mM Tr i s - H C l
8 . 0
4 M MgCl
2
,10mM sodium phosphate
7.0
5 M LiCl, 10 mM sodium phosphate
7.0
denaturation and/or precipitation. When denaturation is used for
elution, the time in which the protein is under harsh conditions
should be as short as possible. Some buffers for denaturation elu-
tion are given in Table 3.8.
References
Amersham Biosciences (2002) Affinity chromatography: principles and
methods. www.chromatography.amershambiosciences.com
Dean PDG, Johnson WS, Middle FA (eds.) (1985) Affinity chromatography:
a practical approach, IRL Press, Oxford
Jack GW (1994) Molec Biotechnol 1:59
Mohr P, Holtzhauer M, Kaiser G (1992) Immunosorption techniques: fun-
damentals and applications. Akademie Verlag, Berlin
Wilchek M, Miron T, Kohn J (1984) Meth Enzymol 104:3
